Triazole compounds that modulate hsp90 activity

ABSTRACT

The present invention relates to substituted triazole compounds and compositions comprising substituted triazole compounds. The invention further relates to methods of inhibiting the activity of Hsp90 in a subject in need thereof and methods for preventing or treating hyperproliferative disorders, such as cancer, in a subject in need thereof comprising administering to the subject a substituted triazole compound of the invention, or a composition comprising such a compound.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.60/964,447, filed on Aug. 13, 2007. The entire teachings of the aboveapplication is incorporated herein by reference.

BACKGROUND OF THE INVENTION

Although tremendous advances have been made in elucidating the genomicabnormalities that cause malignant cancer cells, currently availablechemotherapy remains unsatisfactory, and the prognosis for the majorityof patients diagnosed with cancer remains dismal. Most chemotherapeuticagents act on a specific molecular target thought to be involved in thedevelopment of the malignant phenotype. However, a complex network ofsignaling pathways regulate cell proliferation, and the majority ofmalignant cancers are facilitated by multiple genetic abnormalities inthese pathways. Therefore, it is unlikely that a therapeutic agent thatacts on one molecular target will be fully effective in curing a patientwho has cancer.

Heat shock proteins (HSPs) are a class of chaperone proteins that areup-regulated in response to elevated temperature and other environmentalstresses, such as ultraviolet light, nutrient deprivation, and oxygendeprivation. HSPs act as chaperones to other cellular proteins (calledclient proteins) and facilitate their proper folding and repair, and aidin the refolding of misfolded client proteins. There are several knownfamilies of HSPs, each having its own set of client proteins. The Hsp90family is one of the most abundant HSP families, accounting for about1-2% of proteins in a cell that is not under stress and increasing toabout 4-6% in a cell under stress. Inhibition of Hsp90 results indegradation of its client proteins via the ubiquitin proteasome pathway.Unlike other chaperone proteins, the client proteins of Hsp90 are mostlyprotein kinases or transcription factors involved in signaltransduction, and a number of its client proteins have been shown to beinvolved in the progression of cancer. Examples of Hsp90 client proteinsthat have been implicated in the progression of cancer are describedbelow.

Her-2 is a transmembrane tyrosine kinase cell surface growth factorreceptor that is expressed in normal epithelial cells. Her2 has anextracellular domain that interacts with extracellular growth factorsand an internal tyrosine kinase portion that transmits the externalgrowth signal to the nucleus of the cell. Her2 is overexpressed in asignificant proportion of malignancies, such as breast cancer, ovariancancer, prostate cancer, and gastric cancers, and is typicallyassociated with a poor prognosis.

Akt kinase is a serine/threonine kinase which is a downstream effectormolecule of phosphoinositide 3-kinase and is involved in protecting thecell from apoptosis. Akt kinase is thought to be involved in theprogression of cancer because it stimulates cell proliferation andsuppresses apoptosis.

Cdk4/cyclin D complexes are involved in phosphorylation ofretinoblastoma protein which is an essential step in progression of acell through the G1 phase of the cell cycle. Disruption of Hsp90activity has been shown to decrease the half life of newly synthesizedCdk4.

The Raf family of proto-oncogenes (A-raf, B-raf and C-raf) was firstidentified when C-raf (raf-1) was discovered due to its homology withv-raf, the transforming gene of the mouse sarcoma virus 3611. A-raf waslater discovered by screening a cDNA library under low stringencyconditions using a v-raf probe, and B-raf was discovered due to itshomology with C-Rmil, a transforming gene in avaian retrovirus Mill HillNo. 2. The Raf family of proteins is involved in the Ras/Raf/MEK/ERKpathway, referred to herein as the “MAP kinase pathway” (MEK stands for“MAPK/ERK kinase” and ERK stands for “extracellularly regulatedkinases”), which has been implicated in the genesis and progression ofmany human cancers through upregulation of cell division andproliferation. All raf proteins are serine/theronine kinases which arecapable of activating the MAP kinase pathway. However, B-raf is far morepotent at activating this pathway than A-raf or C-raf, and mutations inthe gene encoding B-raf are more common in cancer. For example, B-rafmutations have been identified in 60% to 70% of malignant melanomas, 83%of anaplastic thyroid carcinoma, 35% to 69% of papillary thyroidcarcinoma, 4% to 16% of colon cancer, 63% of low-grade ovariancarcinoma, 15% of Barrett's esophageal carcinoma, 4% of acute myeloidleukemia, 3-4.8% of head and neck squamous cell carcinoma, 2%-3% ofnon-small-cell lung cancer, 2% of gastric carcinoma, 2% of non-Hodgkinslymphoma and has been reported in glioma, sarcoma, breast cancer,cholangiocarcinoma, and liver cancer. Most mutations in B-raf that havebeen found in human cancers are point mutations that occur in the kinasedomain and are clustered in exons 11 and 15 of the gene which containsseveral regulatory phosphorylation sites (S446, 5447, D448, D449, T599,and S602). (Beeram, et al., Journal of Clinical Oncology (2005),23(27):6771-6790). The most prevalent mutation is the T1799Atransversion mutation which accounts for more than 80% of mutations inthe BRAF gene and results in a V600E mutation in B-raf. The V600E wasformerly designated V599E (the gene mutation was designated T1796A) dueto a mistake in the GenBank nucleotide sequence NM 004333. The correctedGenBank sequence is NT 007914 and designates the protein mutation asV600E and the gene mutation as T1799A. This corrected numbering will beused herein. This mutation is thought to mimic phosphorylation in theactivation segment of B-raf since it inserts a negatively chargedresidue near two activating phosphorylation sites, T599 and S602, andthus results in constitutively active B-raf in a Ras independent manner.(Xing, M., Endocrine-Related Cancer (2005), 12:245-262).

Treatment of cancer cells with 17AAG, an Hsp90 inhibitor, has been shownto stimulate the degradation of B-raf, and mutant forms of B-raf havebeen shown to be more sensitive to degradation than the wild type. Forexample, when melanoma cell line A375 which contain the V600E mutationwas treated with 17AAG, B-raf was degraded more rapidly than in CHLcells which contained wild type B-raf. Other B-raf mutants (e.g., V600D,G469A, G469E, G596R, G466V, and G594V) were a found to be degraded morerapidly than wild type B-raf when transvected into COS cells. However,B-raf mutants E586K and L597V were not sensitive to degradation whencells were treated with 17AAG. Therefore, it is believed that wild typeB-raf in its activated form is a client protein of Hsp90 and that mostmutated forms of B-raf are more dependent on Hsp90 for folding,stability and/or function than the wild type protein. (Dias, et al.,Cancer Res. (2005), 65(23): 10686-10691).

Raf-1 is a MAP 3-kinase (MAP3K) which when activated can phosphorylateand activate the serine/threonine specific protein kinases ERK1 andERK2. Activated ERKs play an important role in the control of geneexpression involved in the cell division cycle, apoptosis, celldifferentiation and cell migration.

Anaplastic large-cell lymphoma (ALCL) is a type of non-Hodgkin'slymphoma characterized by the expression of CD30/Ki-1 antigen. ALCLnormally arises from T-cells, however, a subset of cases have either anull cell or B-cell phenotype. Cases that arise from B-cells aresometimes categorized as diffuse large B-cell lymphomas. About 60% ofthe ALCL case that express CD30/Ki-1 antigen also have the chromosomaltranslocation t(2;5)(p23;q35) which fuses the nucleophosmin (NPM/B23)gene to the anaplastic lymphoma kinase (ALK) gene and results in theoncogenetic fusion protein NPM-ALK with tyrosine kinase activity. Withinspecific subtypes of ALCL, ALK rearrangements have been observed in thefollowing percentages: 1) 30% to 50% of pleomorphic ALCL, 2) more than80% of monomorphic ALCL, 3) 75% to 100% of small-cell cases, and 4) 60%to 100% of lymphohistiocytic ALCL. NPM-ALK is able to transform bothfibroblasts, hematopoietic, and primary bone marrow cell lines, and isthought, to stimulate mitosis through the RAS pathway and the throughactivation of phospholipase C-gamma (PLC-gamma), and to protect againstapoptosis through its activation of phosphatidylinositol 3 kinase (PI-3kinase) survival pathway. (Duyster, et al., Oncogene (2001),20:5623-5637). NPM-ALK has been shown to associate with Hsp90 andincubation of NPM-ALK expressing ALCL cells with the benzoquinoneansamycin, 17AAG, has been shown to disrupt this association resultingin increased degradation of NPM-ALK and induce cell-cycle arrest andapoptosis. (Georgakis, et al., Exp. Hematology (2006), 34(12):1670-1679;Bonvini, et al., Cancer Research (2002), 62:1559-1566).

The transforming protein of Rous sarcoma virus, v-src, is a prototype ofan oncogene family that induces cellular transformation (i.e.,tumorogenesis) by non-regulated kinase activity. Hsp90 has been shown tocomplex with v-scr and inhibit its degradation.

Hsp90 is required to maintain steroid hormone receptors in aconformation capable of binding hormone with high affinity. Inhibitionof the action of Hsp90 therefore is expected to be useful in treatinghormone-associated malignancies such as breast cancer. p53 is a tumorsuppressor protein that causes cell cycle arrest and apoptosis. Mutationof the p53 gene is found in about half of all human cancers making itone of the most common genetic alterations found in cancerous cells. Inaddition, p53 mutation is associated with a poor prognosis. Wild-typep53 has been shown to interact with Hsp90, but mutated p53 forms a morestable association than wild-type p53 as a result of its misfoldedconformations. A stronger interaction with Hsp90 protects the mutatedprotein form normal proteolytic degradation and prolongs its half-life.In a cell that is heterozygous for mutated and wild-type p53, inhibitionof the stabilizing effect of Hsp90 causes mutant p53 to be degraded andrestores the normal transcriptional activity of wild-type p53.

Hif-1α is a hypoxia-inducible transcription factor that is up-regulatedunder low oxygen conditions. Under normal oxygen conditions Hif-1αassociates with Von Hippel-Lindau (VHL) tumor suppressor protein and isdegraded. Low oxygen conditions inhibit this association and allowsHif-1α to accumulate and complex with Hif-1β to form an activetranscription complex that associates with hypoxia-response elements toactivate the transcription of vascular endothelial growth factor (VEGF).Increased Hif-1α is associated with increased metastasis and a poorprognosis.

There are two classes of PKs: protein tyrosine kinases (PTKs), whichcatalyze the phosphorylation of tyrosine kinase residues, and theserine-threonine kinases (STKs), which catalyze the phosphorylation ofserine or threonine residues. Growth factor receptors with PTK activityare known as receptor tyrosine kinases. Receptor tyrosine kinases are afamily of tightly regulated enzymes, and the aberrant activation ofvarious members of the family is one of the hallmarks of cancer. Thereceptor tyrosine kinase family can be divided into subgroups that havesimilar structural organization and sequence similarity within thekinase domain.

Epidermal Growth Factor Receptor (EGFR) is a member of the type 1subgroup of receptor tyrosine kinase family of growth factor receptors,which play critical roles in cellular growth, differentiation, andsurvival. Activation of these receptors typically occurs via specificligand binding which results in hetero- or homodimerization betweenreceptor family members, with subsequent autophosphorylation of thetyrosine kinase domain. Specific ligands which bind to EGFR includeepidermal growth factor (EGF), transforming growth factor α (TGFα,amphiregulin and some viral growth factors. Activation of EGFR triggersa cascade of intracellular signaling pathways involved in both cellularproliferation (the ras/raf/MAP kinase pathway) and survival (the PI3kinase/Akt pathway). Members of this family, including EGFR and HER2,have been directly implicated in cellular transformation.

A number of human malignancies are associated with aberrant oroverexpression of EGFR and/or overexpression of its specific ligands(Gullick, Br. Med. Bull. (1991), 47:87-98; Modijtahedi and Dean, Int. J.Oncol. (1994), 4:277-96; Salomon, et al., Crit. Rev. Oncol. Hematol.(1995);19:183-232, the entire teachings of each of these references areincorporated herein by reference). Aberrant or overexpression of EGFRhas been associated with an adverse prognosis in a number of humancancers, including head and neck, breast, colon, prostate, lung (e.g.,NSCLC, adenocarcinoma and squamous lung cancer), ovaries,gastrointestinal cancers (gastric, colon, pancreatic), renal cellcancer, bladder cancer, glioma, gynecological carcinomas, and prostatecancer. In some instances, overexpression of tumor EGFR has beencorrelated with both chemoresistance and a poor prognosis (Lei, et al.,Anticancer. Res. (1999), 19:221-8; Veale, et al., Br. J. Cancer (1993);68:162-5, the entire teachings of each of these references areincorporated herein by reference).

Gefitinib, a chemotherapeutic agent that inhibits the activity of EGFR,has been found to be highly efficacious in a subset of lung cancerpatients that have mutations in the tyrosine kinase domain of EGFR. Inthe presence of EGF, these mutants displayed two to three times higheractivity than wild type EGFR. In addition, wild type EGFR wasinternalized by the cells and down-regulated after 15 minutes, where asmutant EGFR was internalized more slowly and continued to be activatedfor up to three hours (Lynch, et al., The New England Journal ofMedicine (2006), 350:2129-2139, the entire teachings of which areincorporated herein by reference).

Gliomas are another type of cancer that is characterized byamplification and/or mutation of the EGFR gene. One of the most commonmutations in the EGFR gene is a deletion of exons 2-7 which results in atruncated form of EGFR in which amino acids 6-273 of the extracellulardomain are replaced with a single glycine residue. This mutation iscalled EGFRvIII and is expressed in about half of all glioblastomas.EGFRvIII is unable to bind EGF and TGFα and has constitutive,ligand-independent tyrosine kinase activity. Hsp90 co-purifies withEGFRvIII indicating that Hsp90 complexes with EGFRvIII. Moreover, Hsp90inhibitor geldanamycin, a benzoquinone ansamycin antibiotic, was able todecrease the expression of EGFRvIII indicating that interaction withHsp90 is essential to maintain high expression levels of EGFRvIII(Lavictoire, et al., Journal of Biological Chemistry (2003),278(7):5292-5299, the entire teachings of which are incorporated hereinby reference). These results demonstrate that inhibiting the activity ofHsp90 is an effective strategy for treating cancers that are associatedwith inappropriate EGFR activity.

The members of the type III group of receptor tyrosine kinases includeplatelet-derived growth factor (PDGF) receptors (PDGF receptors alphaand beta), colony-stimulating factor (CSF-1) receptor (CSF-1R, c-Fms),Fms-like tyrosine kinase (FLT3), and stem cell factor receptor (c-kit).FILT3 is primarily expressed on immature hematopoietic progenitors andregulates their proliferation and survival.

Hematologic cancers, also known as hematologic or hematopoieticmalignancies, are cancers of the blood or bone marrow; includingleukemia and lymphoma. Acute myelogenous leukemia (AML) is a clonalhematopoietic stem cell leukemia that represents about 90% of all acuteleukemias in adults with an incidence of 3.9 per 100,000 (See e.g.,Lowenberg et al., N. Eng. J. Med. 341: 1051-62 (1999) and LopesdeMenezes, et al, Clin. Cancer Res. (2005), 11(14):5281-5291, the enterteachings of both references are incorporated by reference). Whilechemotherapy can result in complete remissions, the long termdisease-free survival rate for AML is about 14% with about 7,400 deathsfrom AML each year in the United States. Approximately 70% of AML blastsexpress wild type FLT3 and about 25% to about 35% express FLT3 kinasereceptor mutations which result in constitutively active FLT3. Two typesof activating mutations have been identified in AML patients: internaltandem duplications (ITDs) and point mutation in the activating loop ofthe kinase domain. FLT3-ITD mutations in AML patients is indicative of ap6 or prognosis for survival, and in patients who are in remission,FLT3-ITD mutations are the most significant factor adversely affectingrelapse rate with 64% of patients having the mutation relapsing within 5years (see Current Pharmaceutical Design (2005), 11:3449-3457, theentire teachings of which are incorporated herein by reference). Theprognostic significance of FLT3 mutations in clinical studies suggeststhat FLT3 plays a driving role in AML and may be necessary for thedevelopment and maintenance of the disease.

Mixed Lineage Leukemia (MLL) involve translocations of chromosome 11band q23 (11q23) and occur in approximately 80% of infant hematologicalmalignancies and 10% of adult acute leukemias. Although certain 11q23translocation have been shown to be essential to immortalization ofhematopoietic progenitors in vitro, a secondary genotoxic event isrequired to develop leukemia. There is a strong concordance between FLT3and MLL fusion gene expression, and the most consistently overexpressedgene in MLL is FLT3. Moreover, it has been shown that activated FLT3together with MLL fusion gene expression induces acute leukemia with ashort latency period (see Ono, et al., J. of Clinical Investigation(2005), 115:919-929, the entire teachings of which are incorporated byreference). Therefore, it is believed that FLT3 signally is involved inthe development and maintenance of MLL (see Armstrong, et al., CancerCell (2003), 3:173-183, the entire teachings of which are incorporatedherein by reference).

The FLT3-ITD mutation is also present in about 3% of cases of adultmyelodysplastic syndrome and some cases of acute lymphocytic leukemia(ALL) (Current Pharmaceutical Design (2005), 11:3449-3457).

FLT3 has been shown to be a client protein of Hsp90, and 17AAG, abenzoquinone ansamycin antibiotic that inhibits Hsp90 activity, has beenshown to disrupt the association of Flt3 with Hsp90. The growth ofleukemia cell that express either wild type FLT3 or FLT3-ITD mutationswas found to be inhibited by treatment with 17″AAG (Yao, et al.,Clinical Cancer Research (2003), 9:4483-4493, the entire teachings ofwhich are incorporated herein by reference).

c-Kit is a membrane type III receptor protein tyrosine kinase whichbinds Stem Cell Factor (SCF) to its extracellular domain. c-Kit hastyrosine kinase activity and is required for normal hematopoiesis.However, mutations in c-kit can result in ligand-independent tyrosinekinase activity, autophosphorylation, and uncontrolled cellproliferation. Aberrant expression and/or activation of c-Kit has beenimplicated in a variety of pathologic states. For example, evidence fora contribution of c-Kit to neoplastic pathology includes its associationwith leukemias and mast cell tumors, small cell lung cancer, testicularcancer, and some cancers of the gastrointestinal tract and centralnervous system. In addition, c-Kit has been implicated in playing a rolein carcinogenesis of the female genital tract sarcomas ofneuroectodermal origin, and Schwann cell neoplasia associated withneurofibromatosis. (Yang et al., J Clin Invest. (2003), 112:1851-1861;Viskochil, J Clin Invest. (2003), 112:1791-1793, the entire teachings ofeach of these references are incorporated herein by reference). c-Kithas been shown to be a client protein of Hsp90, and Hsp90 inhibitor17AAG, a benzoquinon ansamycin, has been shown to induce apoptosis inKasumi-1 cells, an acute myeloid leukemia cell line that harbors amutation in c-kit.

c-Met is a receptor tyrosine kinase that is a client protein of Hsp90and is encoded by the Met protooncogene. Hepatocyte growth factor (HGF)(also referred to as scatter factor (SF)) is the natural ligand of c-Metwhich binds to c-Met and leads to a variety of cellular responses suchas proliferation, survival, angiogenesis, wound healing, tissueregeneration, scattering, motility, invasion and branching morphogenesis(Ma et al., Cancer and Metastasis Reviews (2003), 22: 309-325). c-Metand HGF are expressed in numerous tissues, although their expression isnormally confined predominantly to cells of epithelial and mesenchymalorigin, respectively. c-Met and HGF are required for normal mammaliandevelopment and have been shown to be important in cell migration, cellproliferation and survival, morphogenic differentiation, andorganization of 3-dimensional tubular structures (e.g., renal tubularcells, gland formation, etc.). However, dysregulation of c-Met and/orHGF is believed to contribute to tumor growth, dissemination andinvasion in several human cancers. c-Met and/or HGF are highly expressedin numerous cancers and their expression correlates with poor prognosis(Christensen, et al., Cancer Research (2003), 63:7345-7355). Forexample, c-Met receptor mutations have been shown to be expressed in anumber of human cancers including hereditary and sporadic humanpapillary renal carcinomas, ovarian cancer, childhood hepatocellularcarcinoma, metastatic head and neck squamous cell carcinomas, esophagealcancer and gastric cancer. Met gene amplification and over expression ofc-Met has been shown to be associated with both non-small cell lungcancer (NSCLC) and small cell lung cancer (SCLC), as well as colorectalcancer, and the Tpr/Met fusion protein has been shown to be present inhuman osteogenic sarcoma and gastric cancer. Families with germinemutations that activate c-Met kinase are prone to multiple kidney tumorsas well as tumors in other tissues. Numerous studies have correlated theexpression of c-Met and/or HGF with the state of disease progression ofdifferent types of cancer (including lung, colon, breast, prostate,liver, pancreas, brain, kidney, ovarian, stomach, skin, and bonecancers).

The validity of targeting receptor tyrosine kinases (RTK) that aredysregulated in human cancers is illustrated by the successes of Gleevectargeting Bcr-Abl in chronic myelogenous leukemia and c-Kit ingastroinstinal stromal tumors, Herceptin in Her-2 overexpressing breastcancers, and Iressa in select NSCLC that have dysregulated EGFR.Compelling evidence exists for targeting c-Met in the treatment of humancancers and several small drug molecules that inhibit c-Met arecurrently in development. However, therapies that target specific RTKoften work well initially for treating cancer but eventually fail due toadditional mutations which allow RTK to maintain its activity in thepresence of the drug. Moreover, the selective c-Met inhibitor SU11274,while highly affected against wild type c-Met and some mutants of c-Met,has been shown to be ineffective against other c-Met mutants (Berthou,et al., Oncogene (2004), 23:5387-5393). Therefore, a need exists todevelop new anticancer therapeutics that reduce the expression and/oractivity of c-Met via a different mechanism than therapeutics thatdirectly inhibit c-Met.

BCR-ABL is an oncoprotein with tyrosine kinase activity and has beenassociated with chronic myelogenous leukemia (CML), with a subset ofpatients with acute lymphocytic leukemia (ALL) and with a subset ofpatients with acute myelogenous leukemia (AML). In fact, the BCR-ABLoncogene has been found in at least 90-95% of patients with CML, 20% ofadults with ALL, 5% of children with ALL, and in about 2% of adults withAML. The BCR-ABL oncoprotein is generated by the translocation of genesequences from the c-ABL protein tyrosine kinase on chromosome 9 intothe BCR sequences on chromosome 22, producing the Philadelphiachromosome. The BCR-ABL gene has been shown to produce at least threealternative chimeric proteins, p230 Bcr-Abl, p210 Bcr-Abl, and p190Bcr-Abl which have unregulated tyrosine kinase activity. The p210Bcr-Abl fusion protein is most often associated with CML, while the p190Bcr-Abl fusion protein is most often associated with ALL. Bcr-Abl hasalso been associated with a variety of additional hematologicalmalignancies including granulocytic hyperplasia, myelomonocyticleukemia, lymphomas and erythroid leukemia.

Studies have shown that lowering the expression or activity of Bcr-Ablis effective in treating Bcr-Abl-positive leukemias. For example, agentssuch as As₂O₃ which lower Bcr-Abl expression have been shown to behighly effective against Bcr-Abl leukemias. In addition, inhibition ofBcr-Abl tyrosine kinase activity by Imatinib (also known as STI571 andGleevic) induces differentiation and apoptosis and causes eradication ofBcr-Abl positive leukemia cells both in vivo and in vitro. In patientswith CML in the chronic phase, as well as in a blast crisis, treatmentwith Imatinib typically will induce remission. However, in many cases,particularly in those patients who were in a blast crisis beforeremission, the remission is not durable because the Bcr-Abl fusionprotein develops mutations that cause it to be resistance to Imatinib.(See Nimmanapalli, et al., Cancer Research (2001), 61:1799-1804; andGorre, et al., Blood (2002), 100:3041-3044, the entire teachings of eachof these references are incorporated herein by reference).

Bcr-Abl fusion proteins exist as complexes with Hsp90 and are rapidlydegraded when the action of Hsp90 is inhibited. It has been shown thatgeldanamycin, a benzoquinone ansamycin antibiotic that disrupts theassociation of Bcr-Abl with Hsp90, results in proteasomal degradation ofBcr-Abl and induces apoptosis in Bcr-Abl leukemia cells.

Hsp90 has been shown by mutational analysis to be necessary for thesurvival of normal eukaryotic cells. However, Hsp90 is over expressed inmany tumor types indicating that it may play a significant role in thesurvival of cancer cells and that cancer cells may be more sensitive toinhibition of Hsp90 than normal cells. For example, cancer cellstypically have a large number of mutated and overexpressed oncoproteinsthat are dependent on Hsp90 for folding. In addition, because theenvironment of a tumor is typically hostile due to hypoxia, nutrientdeprivation, acidosis, etc., tumor cells may be especially dependent onHsp90 for survival. Moreover, inhibition of Hsp90 causes simultaneousinhibition of a number of oncoproteins, as well as hormone receptors andtranscription factors making it an attractive target for an anti-canceragent. In fact, benzoquinone ansamycins, a family of natural productsthat inhibit Hsp90, has shown evidence of therapeutic activity inclinical trials.

Although promising, benzoquinone ansamycins, and their derivatives,suffer from a number of limitations. For example, they have low oralbioavailability, and their limited solubility makes them difficult toformula. In addition, they are metabolized by polymorphic cytochromeP450 CYP3A4 and are a substrate for P-glycoprotein export pump involvedin the development of multidrug resistance. Therefore, a need exist fornew therapeutics that improve the prognosis of cancer patients and thatreduces or overcomes the limitations of currently used anti-canceragents.

HSPs are highly conserved from microorganisms to mammals. When apathogen invades a host, both the pathogen and the host increase HSPproduction. HSPs appear to play various roles in the infection process.For instance, Hsp90 has been shown to play a role in the pathwaysinvolved in the uptake and/or killing of bacteria in phagocytic cells.Yan, L. et al., Eukaryotic Cell, 567-578;3(3), 2004. Hsp90 has also beenshown to be essential for the uptake of binary actin ADP-ribosylatingtoxins into eukaryotic cells. Haug, G., Infection and Immunity, 12,3066-3068, 2004. Additionally, Hsp90 has been identified as playing arole in viral proliferation in a number of viruses including influenzavirus, vaccinia virus, herpes simplex virus type I, and HIV-1 virus.Momose, F, et al., J. Biol. Chem., 45306-45314, 277(47), 2002; Hung, J.,et al., J. Virology, 1379-1390, 76(3), 2002; Li, Y., et al.,Antimicrobial Agents and Chemotherapy, 867-872, 48(3), 2004; O'Keefe,B., et al., J. Biol. Chem., 279-287, 275(1), 2000.

Opportunistic fungal infections that are resistant to antifungal drugshave become an increasing problem, particularly in immunocompromisedpatients. Hsp90 has been shown to play a role in the evolution of drugresistance in fungi. Cowen, L. et al., Eukaryotic Cell, 2184-2188,5(12), 2006; Cowen, L. et al., Science, 309:2185-2189, 2005.

SUMMARY OF THE INVENTION

The present invention provides compounds which inhibit the activity ofHsp90 and are useful in the treatment of proliferative disorders, suchas cancer.

In one embodiment, the present invention provides compounds representedby structural formula (I):

wherein:

-   -   R₁, R₂ and R₃ are independently —OH, —SH, —NR₇H, —OR₂₆, —SR₂₆,        —O(CH₂)_(m)OH, —O(CH₂)_(m)SH, —O(CH₂)_(m)NR₇H, —S(CH₂)_(m)OH,        —S(CH₂)_(m)SH, —S(CH₂)_(m)NR₇H, —OC(O)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁,        —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇, —OC(O)OR₇,        —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇, —NR₇CH₂C(O)R₇,        —OCH₂C(O)OR₇, —SCH₂C(O)OR₇; —NR₇CH₂C(O)OR₇, —OCH₂C(O)NR₁₀R₁₁,        —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,        —SS(O)_(p)R₇, —S(O)_(p)OR₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁,        —SS(O)_(p)NR₁₀R₁₁, —NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇,        —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇, —OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇,        —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇, —OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁,        —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇, —NR₇C(NR₈)R₇,        —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,        —SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —OP(O)(OR₇)₂, or        —SP(O)(OR₇)₂, provided that at least one of R₁, R₂ and R₃ is        —OP(O)(OH)₂;    -   R₅ is —X₂₀R₅₀, an optionally substituted alkyl, an optionally        substituted alkenyl, an optionally substituted alkynyl, an        optionally substituted cycloalkyl, an optionally substituted        cycloalkenyl, an optionally substituted heterocyclyl, an        optionally substituted aryl, an optionally substituted        heteroaryl, an optionally substituted aralkyl, or an optionally        substituted heteraralkyl;    -   R₇ and R₈, for each occurrence, is independently, —H, an        optionally substituted alkyl, an optionally substituted alkenyl,        an optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;    -   R₁₀ and R₁₁, for each occurrence, is independently —H, an        optionally substituted alkyl, an optionally substituted alkenyl,        an optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;        or R₁₀ and R₁₁, taken together with the nitrogen to which they        are attached, form an optionally substituted heterocyclyl or an        optionally substituted heteroaryl;    -   R₂₆ is a lower alkyl;    -   R₅₀ is an optionally substituted aryl or an optionally        substituted heteroaryl;    -   X₂₀ is a C1-C4 alkyl, NR₇, C(O), C(S), C(NR₈), or S(O)_(p);    -   Z is a substituent;    -   p, for each occurrence, is independently, 1 or 2;    -   m for each occurrence, is independently 1, 2, 3, or 4; and    -   n is 0, 1, 2, or 3;    -   or a tautomer, pharmaceutically acceptable salt, solvate,        clathrate or a prodrug thereof.

In one embodiment, of the compounds represented by formula (I), thecompound is not3-hydroxy-4-(5-mercapto-4-(naphthalen-1-yl)-4H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate.

The compounds shown in Table 1 or compounds of any formula herein, ortautomers, pharmaceutically acceptable salts, solvates, clathrates,hydrates, polymorphs or prodrugs thereof, inhibit the activity of Hsp90and, thereby facilitates the degradation of Hsp90 client proteins. Hsp90is necessary for the survival of normal eukaryotic cells. However, Hsp90is over expressed in many tumor types indicating that it may play asignificant role in the survival of cancer cells and that cancer cellsmay be more sensitive to inhibition of Hsp90 than normal cells. Thus,the compounds shown in Table 1 or compounds of any formula herein, ortautomers, pharmaceutically acceptable salts, solvates, clathrates,hydrates, polymorphs or prodrugs thereof, are useful treatingproliferative disorders such as cancer.

Although chemotherapeutic agents initially cause tumor regression, mostagents that are currently used to treat cancer target only one pathwayto tumor progression. Therefore, in many instances, after treatment withone or more chemotherapeutic agents, a tumor develops multidrugresistance and no longer responses positively to treatment. One of theadvantages of inhibiting Hsp90 activity is that several of its clientproteins, which are mostly protein kinases or transcription factorsinvolved in signal transduction, have been shown to be involved in theprogression of cancer. Thus, inhibition of Hsp90 provides a method ofshort circuiting several pathways for tumor progression simultaneously.Therefore, treatment of tumors with an Hsp90 inhibitor of the inventioneither alone, or in combination with other chemotherapeutic agents, ismore likely to result in regression or elimination of the tumor, andless likely to result in the development of more aggressive multidrugresistant tumors than other currently available therapies.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a SCID mouse xenograft study to determine the effects ofCompound #3 on the in vivo growth rate of the human multiple myelomatumor cell line RPMI 8226.

FIG. 2 is a SCID mouse xenograft study to determine the effects ofCompound #3 on the in vivo growth rate of the canine osteosarcoma tumorcell line D17.

DETAILED DESCRIPTION OF THE INVENTION

A description of preferred embodiments of the invention follows.

The present invention provides compounds disclosed herein and uses ofsaid compounds to inhibit Hsp90 activity and for the treatment of aproliferative disorder, such as cancer. In particular, the presentinvention encompasses the use of compounds of the invention to slow orstop the growth of cancerous cells or to reduce or eliminate cancerouscells in a subject, preferably the subject is a mammal.

In certain embodiments, the compounds of the invention can be used incombination with other chemotherapeutic agents and may help to preventor reduce the development of multidrug resistant cancerous cells in amammal. In this embodiment, the compounds of the invention may allow areduced efficacious amount of a second chemotherapeutic agent given to amammal, because compounds of the invention should inhibit thedevelopment of multidrug resistant cancerous cells.

In certain embodiments, the compounds of the invention can be used toblock, occlude, or otherwise disrupt blood flow in neovasculature.

In other embodiments, the compounds of the invention can be used totreat or inhibit angiogenesis in a subject in need thereof.

The present invention also relates to compounds which inhibit theactivity of topoisomerase II.

The present invention also relates to the discovery that treatment ofcells, such as peripheral blood mononuclear cells (PMBCs) that have beenstimulated with an inflammatory stimulus, such as INFγ/LPS or SAC, withan Hsp90 inhibitor reduces the expression of GR in the PMBCs and reducesthe production of inflammatory cytokines.

The present invention also relates to compounds which inhibit theactivity of Hsp90 and are useful in the treatment of or prevention ofinfections.

In another embodiment, the present invention relates to a method oftreating or preventing fungal drug resistance in a mammal in need ofsuch treatment. The method comprises administering to the mammal aneffective amount of an Hsp90 inhibitor disclosed herein.

In another embodiment, the present invention relates to methods ofadministering a dosage solution of compounds of the present invention toa mammal.

A. Terminology

Unless otherwise specified, the below terms used herein are defined asfollows:

As used herein, the term “alkyl” means a saturated straight chain orbranched non-cyclic hydrocarbon having from 1 to 10 carbon atoms.Representative saturated straight chain alkyls include methyl, ethyl,n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl andn-decyl; while saturated branched alkyls include isopropyl, sec-butyl,isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl,2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl,3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl,2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl,2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl,2,2-dimethylhexyl, 3,3-dimethylpentyl, 3,3-dimethylhexyl,4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl, 2-ethylhexyl,3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl,2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl,2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl,3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like. Theterm “(C₁-C₆)alkyl” means a saturated straight chain or branchednon-cyclic hydrocarbon having from 1 to 6 carbon atoms. Representative(C₁-C₆)alkyl groups are those shown above having from 1 to 6 carbonatoms. Alkyl groups included in compounds of this invention may beoptionally substituted with one or more substituents.

As used herein, the term “alkenyl” means a saturated straight chain orbranched non-cyclic hydrocarbon having from 2 to 10 carbon atoms andhaving at least one carbon-carbon double bond. Representative straightchain and branched (C₂-C₁₀)alkenyls include vinyl, allyl, 1-butenyl,2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl,3-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl,3-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 2-decenyl,3-decenyl and the like. Alkenyl groups may be optionally substitutedwith one or more substituents.

As used herein, the term “alkynyl” means a saturated straight chain orbranched non-cyclic hydrocarbon having from 2 to 10 carbon atoms andhaving at least one carbon-carbon triple bond. Representative straightchain and branched alkynyls include acetylenyl, propynyl, 1-butynyl,2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, 4-pentynyl,1-hexynyl, 2-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 6-heptynyl,1-octynyl, 2-octynyl, 7-octynyl, 1-nonynyl, 2-nonynyl, 8-nonynyl,1-decynyl, 2-decynyl, 9-decynyl, and the like. Alkynyl groups may beoptionally substituted with one or more substituents.

As used herein, the term “cycloalkyl” means a saturated, mono- orpolycyclic alkyl radical having from 3 to 20 carbon atoms.Representative cycloalkyls include cyclopropyl, 1-methylcyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,cyclononyl, -cyclodecyl, octahydro-pentalenyl, and the like. Cycloalkylgroups may be optionally substituted with one or more substituents.

As used herein, the term “cycloalkenyl” means a mono- or poly-cyclicnon-aromatic alkyl radical having at least one carbon-carbon double bondin the cyclic system and from 3 to 20 carbon atoms. Representativecycloalkenyls include cyclopentenyl, cyclopentadienyl, cyclohexenyl,cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptatrienyl,cyclooctenyl, cyclooctadienyl, cyclooctatrienyl, cyclooctatetraenyl,cyclononenyl, cyclononadienyl, cyclodecenyl, cyclodecadienyl,1,2,3,4,5,8-hexahydronaphthalenyl and the like. Cycloalkenyl groups maybe optionally substituted with one or more substituents.

As used herein, the term “haloalkyl” means an alkyl group, in which oneor more (including all) the hydrogen radicals are replaced by a halogroup, wherein each halo group is independently selected from —F, —Cl,—Br, and —I. The term “halomethyl” means a methyl in which one to threehydrogen radical(s) have been replaced by a halo group. Representativehaloalkyl groups include trifluoromethyl, bromomethyl,1,2-dichloroethyl, 4-iodobutyl, 2-fluoropentyl, and the like.

As used herein, an “alkoxy” is an alkyl group which is attached toanother moiety via an oxygen linker.

As used herein, an “haloalkoxy” is an haloalkyl group which is attachedto another moiety via an oxygen linker.

As used herein, the term an “aromatic ring” or “aryl” means ahydrocarbon monocyclic or polycyclic radical in which at least one ringis aromatic. Examples of suitable aryl groups include, but are notlimited to, phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl,and naphthyl, as well as benzo-fused carbocyclic moieties such as5,6,7,8-tetrahydronaphthyl. Aryl groups may be optionally substitutedwith one or more substituents. In one embodiment, the aryl group is amonocyclic ring, wherein the ring comprises 6 carbon atoms, referred toherein as “(C₆)aryl.”

As used herein, the term “aralkyl” means an aryl group that is attachedto another group by a (C₁-C₆)alkylene group. Representative aralkylgroups include benzyl, 2-phenyl-ethyl, naphth-3-yl-methyl and the like.Aralkyl groups may be optionally substituted with one or moresubstituents.

As used herein, the term “alkylene” refers to an alkyl group that hastwo points of attachment. The term “(C₁-C₆)alkylene” refers to analkylene group that has from one to six carbon atoms. Straight chain(C₁-C₆)alkylene groups are preferred. Non-limiting examples of alkylenegroups include methylene (—CH₂—), ethylene (—CH₂CH₂—), n-propylene(—CH₂CH₂CH₂—), isopropylene (—CH₂CH(CH₃)—), and the like. Alkylenegroups may be optionally substituted with one or more substituents.

As used herein, the term “heterocyclyl” means a monocyclic (typicallyhaving 3- to 10-members) or a polycyclic (typically having 7- to20-members) heterocyclic ring system which is either a saturated ring ora unsaturated non-aromatic ring. A 3- to 10-membered heterocycle cancontain up to 5 heteroatoms; and a 7- to 20-membered heterocycle cancontain up to 7 heteroatoms. Typically, a heterocycle has at least onecarbon atom ring member. Each heteroatom is independently selected fromnitrogen, which can be oxidized (e.g., N(O)) or quaternized; oxygen; andsulfur, including sulfoxide and sulfone. The heterocycle may be attachedvia any heteroatom or carbon atom. Representative heterocycles includemorpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl,tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, andthe like. A heteroatom may be substituted with a protecting group knownto those of ordinary skill in the art, for example, the hydrogen on anitrogen may be substituted with a tert-butoxycarbonyl group.Furthermore, the heterocyclyl may be optionally substituted with one ormore substituents. Only stable isomers of such substituted heterocyclicgroups are contemplated in this definition.

As used herein, the term “heteroaromatic”, “heteroaryl” or like termsmeans a monocyclic or polycyclic heteroaromatic ring comprising carbonatom ring members and one or more heteroatom ring members. Eachheteroatom is independently selected from nitrogen, which can beoxidized (e.g., N(O)) or quaternized; oxygen; and sulfur, includingsulfoxide and sulfone. Representative heteroaryl groups include pyridyl,1-oxo-pyridyl, furanyl, benzo[1,3]dioxolyl, benzo[1,4]dioxinyl, thienyl,pyrrolyl, oxazolyl, imidazolyl, thiazolyl, a isoxazolyl, quinolinyl,pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, atriazinyl, triazolyl, thiadiazolyl, isoquinolinyl, indazolyl,benzoxazolyl, benzofuryl, indolizinyl, imidazopyridyl, tetrazolyl,benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl,indolyl, tetrahydroindolyl, azaindolyl, imidazopyridyl, quinazolinyl,purinyl, pyrrolo[2,3]pyrimidinyl, pyrazolo[3,4]pyrimidinyl,imidazo[1,2-a]pyridyl, and benzothienyl. In one embodiment, theheteroaromatic ring is selected from 5-8 membered monocyclic heteroarylrings. The point of attachment of a heteroaromatic or heteroaryl ringmay be at either a carbon atom or a heteroatom of the heteroaromatic orheteroaryl rings. Heteroaryl groups may be optionally substituted withone or more substituents.

As used herein, the term “(C₅)heteroaryl” means an aromatic heterocyclicring of 5 members, wherein at least one carbon atom of the ring isreplaced with a heteroatom such as, for example, oxygen, sulfur ornitrogen. Representative (C₅)heteroaryls include furanyl, thienyl,pyrrolyl, oxazolyl, imidazolyl, thiazolyl, isoxazolyl, pyrazolyl,isothiazolyl, pyrazinyl, triazolyl, thiadiazolyl, and the like.

As used herein, the term “(C₆)heteroaryl” means an aromatic heterocyclicring of 6 members, wherein at least one carbon atom of the ring isreplaced with a heteroatom such as, for example, oxygen, nitrogen orsulfur. Representative (C₆)heteroaryls include pyridyl, pyridazinyl,pyrazinyl, triazinyl, tetrazinyl and the like.

As used herein, the term “heteroaralkyl” means a heteroaryl group thatis attached to another group by a (C₁-C₆)alkylene. Representativeheteroaralkyls include 2-(pyridin-4-yl)-propyl, 2-(thien-3-yl)-ethyl,imidazol-4-yl-methyl and the like. Heteroaralkyl groups may beoptionally substituted with one or more substituents.

As used herein, the term “halogen” or “halo” means —F, —Cl, —Br or —I.

As used herein the term “heteroalkyl” means a linear straight orbranched chain alkyl group, wherein one or more of the internal carbonatoms in the chain is replaced by a heteroatom, such as, O, N or S,e.g., —[CH₂]_(x)—O—[CH₂]_(y)[CH₃] wherein x is a positive integer and yis 0 or a positive integer, and wherein replacement of the carbon atomdoes not result in a unstable compound.

Suitable substituents for an alkyl, alkylene, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, heterocyclyl, aryl, aralkyl, heteroaryl, andheteroaralkyl groups include are those substituents which form a stablecompound of the invention without significantly adversely affecting thereactivity or biological activity of the compound of the invention.Examples of substituents for an alkyl, alkylene, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, heterocyclyl, aryl, aralkyl, heteroaryl, andheteroarylalkyl include an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl, an optionally substituted aryl, anoptionally substituted heteroaryl, an optionally substituted aralkyl, anoptionally substituted heteraralkyl, an optionally substitutedhaloalkyl, an optionally substituted heteroalkyl, optionally substitutedalkoxy, —C(O)NR₂₈R₂₉, —C(S)NR₂₈R₂₉, —C(NR₃₂)NR₂₈R₂₉, —NR₃₃C(O)R₃₁,—NR₃₃C(S)R₃₁, —NR₃₃C(NR₃₂)R₃₁, halo, —OR₃₃, cyano, nitro, haloalkoxy,—C(O)R₃₃, —C(S)R₃₃, —C(NR₃₂)R₃₃, —NR₂₈R₂₉, —C(O)OR₃₃, —C(S)OR₃₃,—C(NR₃₂)OR₃₃, —OC(O)R₃₃, —OC(S)R₃₃, —OC(NR₃₂)R₃₃, —NR₃₀C(O)NR₂₈R₂₉,—NR₃₃C(S)NR₂₈R₂₉, —NR₃₃C(NR₃₂)NR₂₈R₂₉, —OC(O)NR₂₈NR₂₉, —OC(S)NR₂₈R₂₉,—OC(NR₃₂)NR₂₈R₂₉, —NR₃₃C(O)OR₃₁, —NR₃₃C(S)OR₃₁, —NR₃₃C(NR₃₂)OR₃₁,—S(O)_(h)R₃₃, —OS(O)_(p)R₃₃, —NR₃₃S(O)_(p)R₃₃, —S(O)_(p)NR₂₈R₂₉,—OS(O)_(p)NR₂₈R₂₉, or —NR₃₃S(O)_(p)NR₂₈R₂₉ guanadino, —C(O)SR₃₁,—C(S)SR₃₁, —C(NR₃₂)SR₃₁, —OC(O)OR₃₁, —OC(S)OR₃₁, —OC(NR₃₂)OR₃₁,—SC(O)R₃₃, —SC(O)OR₃₁, —SC(NR₃₂)OR₃₁, —SC(S)R₃₃, —SC(S)OR₃₁,—SC(O)NR₂₈R₂₉, —SC(NR₃₂)NR₂₈R₂₈, —SC(S)NR₂₈R₂₉, —SC(NR₃₂)R₃₃,—OS(O)_(p)OR₃₁, —S(O)_(p)OR₃₁, —NR₃₀S(O)_(p)OR₃₁, —SS(O)_(p)R₃₃,—SS(O)_(p)OR₃₁, —SS(O)_(p)NR₂₈R₂₉, —OP(O)(OR₃₁)₂, or —SP(O)(OR₃₁)₂,(preferably the alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,heterocyclyl, aryl, heteroaryl, aralkyl, heteroalkyl, alkoxy,heteroaralkyl and haloalkyl are unsubstituted); wherein R₂₈ and R₂₉, foreach occurrence is independently, H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl(preferably the alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,heterocyclyl, aryl, heteroaryl, aralkyl and heteraralkyl areunsubstituted);

R₃₃ and R₃₁ for each occurrence is independently, H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl (preferably the alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, heterocyclyl, aryl, heteroaryl, aralkyl, and heteraralkylare unsubstituted); and

R₃₂, for each occurrence is independently, H, an optionally substitutedalkyl, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedCycloalkenyl; an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, an optionally substituted heteraralkyl, —C(O)R₃₃,—C(O)NR₂₈R₂₉, —S(O)_(p)R₃₃, or —S(O)_(p)NR₂₈R₂₉ (preferably the alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl,heteroaryl, aralkyl and heteraralkyl are unsubstituted);

-   -   p is 0, 1 or 2; and    -   h is 0, 1 or 2.

In addition, alkyl, cycloalkyl, alkylene, a heterocyclyl, and anysaturated portion of a alkenyl, cycloalkenyl, alkynyl, aralkyl, andheteroaralkyl groups, may also be substituted with ═O, ═S, ═N—R₃₂.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

As used herein, the terms “subject”, “patient” and “mammal” are usedinterchangeably. The terms “subject” and “patient” refer to an animal(e.g., a bird such as a chicken, quail or turkey, or a mammal),preferably a mammal including a non-primate (e.g., a cow, pig, horse,sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and a primate(e.g., a monkey, chimpanzee and a human), and more preferably a human.In one embodiment, the subject is a non-human animal such as a farmanimal (e.g., a horse, cow, pig or sheep), or a pet (e.g., a dog, cat,guinea pig or rabbit). In a preferred embodiment, the subject is ahuman.

As used herein, the term “lower” refers to a group having up to fouratoms. For example, a “lower alkyl” refers to an alkyl radical havingfrom 1 to 4 carbon atoms, “lower alkoxy” refers to “—O—(C₁-C₄)alkyl anda “lower alkenyl” or “lower alkynyl” refers to an alkenyl or alkynylradical having from 2 to 4 carbon atoms, respectively.

Unless indicated otherwise, the compounds of the invention containingreactive functional groups (such as (without limitation) carboxy,hydroxy, thiol, and amino moieties) also include protected derivativesthereof “Protected derivatives” are those compounds in which a reactivesite or sites are blocked with one or more protecting groups. Examplesof suitable protecting groups for hydroxyl groups include benzyl,methoxymethyl, allyl, trimethylsilyl, tert-butyldimethylsilyl, acetate,and the like. Examples of suitable amine protecting groups includebenzyloxycarbonyl, tert-butoxycarbonyl, tert-butyl, benzyl andfluorenylmethyloxy-carbonyl (Fmoc). Examples of suitable thiolprotecting groups include benzyl, tert-butyl, acetyl, methoxymethyl andthe like. Other suitable protecting groups are well known to those ofordinary skill in the art and include those found in T. W. Greene,Protecting Groups in Organic Synthesis, John Wiley & Sons, Inc. 1981.

As used herein, the term “compound(s) of this invention” and similarterms refers to a compound of formula (I)-(VIII), or Table 1, or apharmaceutically acceptable salt, solvate, clathrate, hydrate, polymorphor prodrug thereof, and also include protected derivatives thereof.

The compounds of the invention may contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers, ordiastereomers. According to this invention, the chemical structuresdepicted herein, including the compounds of this invention, encompassall of the corresponding compounds' enantiomers, diastereomers andgeometric isomers, that is, both the stereochemically pure form (e.g.,geometrically pure, enantiomerically pure, or diastereomerically pure)and isomeric mixtures (e.g., enantiomeric, diastereomeric and geometricisomeric mixtures). In some cases, one enantiomer, diastereomer orgeometric isomer will possess superior activity or an improved toxicityor kinetic profile compared to other isomers. In those cases, suchenantiomers, diastereomers and geometric isomers of compounds of thisinvention are preferred.

As used herein, the term “polymorph” means solid crystalline forms of acompound of the present invention or complex thereof. Differentpolymorphs of the same compound can exhibit different physical, chemicaland/or spectroscopic properties. Different physical properties include,but are not limited to stability (e.g., to heat or light),compressibility and density (important in formulation and productmanufacturing), and dissolution rates (which can affectbioavailability). Differences in stability can result from changes inchemical reactivity (e.g., differential oxidation, such that a dosageform discolors more rapidly when comprised of one polymorph than whencomprised of another polymorph) or mechanical characteristics (e.g.,tablets crumble on storage as a kinetically favored polymorph convertsto thermodynamically more stable polymorph) or both (e.g., tablets ofone polymorph are more susceptible to breakdown at high humidity).Different physical properties of polymorphs can affect their processing.For example, one polymorph might be more likely to form solvates ormight be more difficult to filter or wash free of impurities thananother due to, for example, the shape or size distribution of particlesof it.

As used herein, the term “hydrate” means a compound of the presentinvention or a salt thereof, that further includes a stoichiometric ornon-stoichiometric amount of water bound by non-covalent intermolecularforces.

As used herein, the term “clathrate” means a compound of the presentinvention or a salt thereof in the form of a crystal lattice thatcontains spaces (e.g., channels) that have a guest molecule (e.g., asolvent or water) trapped within.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound of this invention. Prodrugs may become active upon suchreaction under biological conditions, or they may have activity in theirunreacted forms. Examples of prodrugs contemplated in this inventioninclude, but are not limited to, analogs or derivatives of compounds offormula (I)-(VIII), or Table 1 that comprise biohydrolyzable moietiessuch as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzablecarbamates, biohydrolyzable carbonates, biohydrolyzable ureides, andbiohydrolyzable phosphate analogues. Other examples of prodrugs includederivatives of compounds of formula (I)-(VIII), or Table 1 that comprise—NO, —NO₂, —ONO, or —ONO₂ moieties. Prodrugs can typically be preparedusing well-known methods, such as those described by 1 BURGER'SMEDICINAL CHEMISTRY AND DRUG DISCOVERY (1995) 172-178, 949-982 (ManfredE. Wolff ed., 5^(th) ed).

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide”, “biohydrolyzable ester”, “biohydrolyzablecarbamate”, “biohydrolyzable carbonate”, “biohydrolyzable ureide” and“biohydrolyzable phosphate analogue” mean an amide, ester, carbamate,carbonate, ureide, or phosphate analogue, respectively, that either: 1)does not destroy the biological activity of the compound and confersupon that compound advantageous properties in vivo, such as improvedwater solubility, improved circulating half-life in the blood (e.g.,because of reduced metabolism of the prodrug), improved uptake, improvedduration of action, or improved onset of action; or 2) is itselfbiologically inactive but is converted in vivo to a biologically activecompound. Examples of biohydrolyzable amides include, but are notlimited to, lower alkyl amides, α-amino acid amides, alkoxyacyl amides,and alkylaminoalkylcarbonyl amides. Examples of biohydrolyzable estersinclude, but are not limited to, lower alkyl esters, alkoxyacyloxyesters, alkyl acylamino alkyl esters, and choline esters. Examples ofbiohydrolyzable carbamates include, but are not limited to, loweralkylamines, substituted ethylenediamines, aminoacids,hydroxyalkylamines, heterocyclic and heteroaromatic amines, andpolyether amines.

As used herein, “Hsp90” includes each member of the family of heat shockproteins having a mass of about 90-kiloDaltons. For example, in humansthe highly conserved Hsp90 family includes cytosolic Hsp90α and Hsp90βisoforms, as well as GRP94, which is found in the endoplasmic reticulum,and HSP75/TRAP1, which is found in the mitochondrial matrix.

c-Met is a receptor tyrosine kinase that is expressed in normal andmalignant cells and has been identified as a proto-oncogene. HGF/c-Metsignaling triggers an invasive growth program that is thought to beessential in early embryonic development but when dysregulated canresult malignant growth, motility, migration and invasion by a mechanismthat is not yet completely understood. The human Met gene is located onchromosome 7 band 7q21-q31 and spans more than 120 kb (Ma, et al.,Cancer and Metastasis Reviews (2003), 22:309-325). In wild type cells,c-Met is a heterodimer that consists of an extracellular α-subunit and an-subunit with a large extracellular domain, a membrane spanning segmentand an intracellular tyrosine kinase domain. Functional structures anddomains of c-Met include 1) Sema domain at the N-terminus which includesa MRS cysteine-rich region; 2) PSI domain which is also found inplexins, semaphorins and integrins; 3) IPT repeats which are found inimmunoglobulin, plexins and transcription factors; 4) transmembranedomain; 5) juxtamembrane domain; and 6) the intracellular tyrosinekinase domain at the C-terminus.

Activation of c-Met signaling is dependent on phosphorylation ofmultiple residues on c-Met. Upon binding of HGF, c-Met undergoesautophosphorylation at Y1230, Y1234, and Y1235 in the activation loop ofthe tyrosine kinase domain which activates the kinase activity of c-Met.Y1313 can also be phosphorylated in response to HGF binding and isimportant in binding PI3-K which is implicated in prosurvival signaling.Phosphorylation of Y1349 and Y1356 at the C-terminus of c-Met activatesthe multisubstrate signal transducer docking site which has beenimplicated in Met-mediated signal transduction and mediates theinteractions of SHC, Src, and Gab1, while recruitment of Grb2, PI3-K,PLC-γ and SHP2 is dependent on phosphorylation of Y1356 alone.Regulation of cell morphogenesis is mediated via Y1365. Phosphorylationof the Y1003 residue in the juxtamembrane domain mediates the binding ofc-Cbl. c-Cbl acts as a negative regulator protein of c-Met by promotingthe polyubiquitinization of c-Met which leads to degradation.

Dysregulation of HGF/c-Met signaling can be caused by 1) increasedexpression of HGF; 2) activating mutations which typically occur in thetyrosine kinase domain or the juxtamembrane domain of c-Met and conferconstitutive kinase activity; 3) intra-chromosomal amplification of theMet gene and over expression of c-Met; 4) chromosomal translocation suchas in the Trp/Met fusion protein which results in the loss of thejuxtamembrane domain and leads to constitutive activation; and 5)alternate splicing variants c-Met mRNA which lead to loss of thejuxtamembrane domain and also lead to constitutive activation.

Activating mutation in the tyrosine kinase domain or in thejuxtamembrane domain of c-Met which result in increased activation ofthe tyrosine kinase activity have been observed in hereditary andsporatic papillary renal carcinoma, ovarian cancer, hepatocellularcarcinoma, metastatic head and neck squamous cell carcinomas, NSCLC,SCLC, glioma, breast cancer, and gastric cancer. In somatic papillaryrenal cell carcinoma activating mutations have been found at amino acidresidues M1268 (e.g., M1268T), Y1248 (e.g., Y1248D, Y1248H), Y1246(e.g., Y1246H), Y1230 (e.g., Y1230C), L1213 (e.g., L1213V), H1124 (e.g.,H1124D, H1112L, and H1112Y), and V1110 (V1110I). In germline papillaryrenal cell carcinoma activating mutations have been found at amino acidresidues Y1248 (e.g., Y1248C), Y1246 (e.g., Y1246N), V1238 (e.g.,V12381), Y1230 (e.g., Y1230C and Y1230H), V1206 (e.g., V1206L), M1149(e.g., M1149T), and H1112 (e.g., H1112R). In hepatocellular carcinomaactivating mutations have been found at amino acid residues M1268 (e.g.,M12681), K1262 (e.g., K1262R), and T1191 (e.g., T1191I). In head andneck squamous cell carcinoma activating mutations have been found atamino acid residues Y1253 (e.g., Y1253D), Y1235 (e.g., Y1235D), andY1230 (e.g., Y1230C and Y1230D). In glioma activating mutations havebeen found at amino acid residue G1137 (e.g., G1137V). In NSCLCactivating mutations have been found at amino acid residue T1010 (e.g.,T1010I). In SCLC activating mutations have been found at amino acidresidues R988 (e.g., R988C) and T1010 (e.g., T1010I). In breast canceractivating mutations have been found at amino acid residues T1010 (e.g.,T10100. In gastric cancer activating mutations have been found at aminoacid residue P1009 (e.g., P1009S). Amino acids listed herein for c-Metare numbered as in Schmit, et al., Onogene (1999), 18:2343-2350.Compounds of the invention cause the degradation of c-Met and can beused either alone or in combination with other anticancer therapies totreat patients with cancers that have activating mutations in thetyrosine kinase domain or in the 5, juxtamembrane domain of c-Met.

The juxtamembrane of receptor tyrosine kinases has been shown to represscatalytic function and mutation in the juxtamembrane relieve thisrepression and can lead to oncogenesis. The Tpr/Met fusion proteinresults from replacement of the 5′ region of the Met gene with Tpr whichprovides two strong dimerization motifs. Dimerization activates the Metkinase activity and results in transforming and metastatic properties.The Tpr/Met fusion protein has been found in gastric cancer and resultsin increased Met kinase activity. In addition, an alternative splicingform of Met mRNA has been found in small cell lung cancer which resultsin skipping the juxtamembrane domain. Loss of the juxtamembrane domainleads to increased Met kinase activity and oncogenesis. Compounds of theinvention cause the degradation of c-Met and can be used either alone orin combination with other anticancer therapies to treat patients withcancers that have juxtamembrane mutations or deletions in c-Met.

Amplification of the Met gene and overexpression of c-Met has been foundin several types of cancers including gastric cancer, esophageal cancer,small cell lung cancer, non-small cell lung cancer, breast cancer,multiple myeloma, and colorectal cancer metastases. Compounds of theinvention cause the degradation of c-Met and can be used either alone orin combination with other anticancer therapies to treat patients withcancers that have Met gene amplification and/or c-Met overexpression.

Met amplification and mutation has also been implicated as a strategy bywhich certain cancers become resistant to therapy (e.g., chemotherapy orradiation therapy). For example, certain non-small cell lung cancerscontain an activating mutation in receptor tyrosine kinase EGFR whichresults in oncogenesis. Most EGFR mutant NSCLCs initially respond toEGFR inhibitors such as Iressa and Tarceva but the vast majority ofthese tumors ultimately become resistant to the drug. A subset of theseresistant cancers have been shown to have amplified Met, and it isthought that Met amplification is a mechanisms of acquired resistance,and in particular acquired resistance to kinase inhibitors such asIressa and Tarceva (EGFR inhibitors), Gleevec (a Bcr-Abl, PDGF, andc-Kit inhibitor) (Engelman et al., Sciencexpress,www.sciencexpress.org/26 Apr. 2007/page 1/10.1126/science. 1141478).Compounds of the invention cause the degradation of c-Met and can beused either alone or in combination with other anticancer therapies,such as treatment with kinase inhibitors, to treat patients with cancerthat has become resistant to other anticancer therapies.

As used herein, “c-Met associated cancer” refers to any type ofmalignant growth or metastasis that is caused by or enhanced bydysregulation in HGF/c-Met signaling.

B-raf is a serine/threonine kinase that is involved in the MAP kinasepathway and is encoded by a gene located on chromosome 7q32. Tenisoforms of B-raf have been identified which are the result of splicingvariants. The term “B-raf” refers to all such splicing variants. B-rafhas three conserved regions (CR): 1) CR1 which contains a cysteine richdomain (CRD) and most of the Ras binding domain (RBD) and facilitatesthe binding of B-raf to Ras and recruitment to the cell membrane; 2) CR2which is rich in serine and threonine and includes the S365 residuewhich is an inhibitory phosphorylation site; and 3) CR3 which containsthe kinase domain including a G-loop GXGXXG motif, an activation segmentand regulatory phosphorylation sites S446, S447, D448, D449, T599 andS602. B-raf is translocated to the cell membrane and activated byassociation with GTP-bound Ras. B-raf is regulated by changes in itsconformation and is inactive when the activation segment is held in aninactive conformation as a result of hydrophobic interactions with theP-loop. Phosphorylation in the activation segment results in a shift tothe active conformation of B-raf. Interestingly, the activation segmentand P-loop that interact with one-another and restraining the activationsegment in an inactive conformation, are where the majority of B-rafoncogenic mutations are clustered. This indicates that as a result ofB-raf mutations the inactive B-raf conformation is destabilized therebypromoting an active B-raf conformation. (Berram, et al., Journal ofClinical Oncology (2005), 23(27):6771-6790).

B-raf associated cancers are cancers in which inappropriate B-rafactivity is detected. In one embodiment, B-raf associated cancers haveincreased B-raf activity, such as B-raf with mutations in the kinasedomain that confer increased activity over that of wild type B-rafand/or constitutively active B-raf (e.g., B-raf that has activity thatis not dependent on interaction with Ras). Activating mutations in thekinase domain include V600E, V600D, G596R, G594V, G469A, G469E, G466V,and G464V mutations. Examples of B-raf associated cancers includemalignant melanomas, anaplastic thyroid carcinoma, papillary thyroidcarcinoma, follicular thyroid cancer, para-follicular C-cell-derivedmedullary thyroid cancer, colon cancer, ovarian carcinoma, Barrett'sesophageal carcinoma, acute myeloid leukemia, head and neck squamouscell carcinoma, non-small-cell lung cancer, gastric carcinoma,non-Hodgkins lymphoma, glioma, sarcoma, breast cancer,cholangiocarcinoma, and liver cancer in which inappropriate B-rafactivity can be detected, such as increased B-raf activity of a mutantform of B-raf over that of wild type B-raf or constitutive activity ofB-raf.

As used herein, “NPM-ALK” refers to a fusion protein which is the resultof a t(2;5)(p23;q35) translocation of the gene sequence for NPM/B23nucleolar protein into the sequence which encodes for the tyrosinekinase ALK. Typically, the NPM-ALK fusion protein contains the first 117amino acids of the amine terminal of NPM and the C-terminal residues1058 to 1620 of ALK. For a schematic representation of NPM-ALK see FIG.1 of Duyster, et al., Oncogene (2001), 20:5623-5637, the entireteachings of which are incorporated herein by reference.

The term “NPM-ALK associated cancers” refers to cancers in which theNPM-ALK fusion protein is expressed, such as ALCL and diffuse largeB-cell lymphomas.

The term “c-kit” or “c-kit kinase” refers to a membrane receptor proteintyrosine kinase which is preferably activated upon binding Stem CellFactor (SCF) to its extracellular domain (Yarden et al., 1987; Qiu etal., 1988). The full length amino acid sequence of a c-kit kinasepreferably is as set forth in Yarden, et al., 1987, EMBO J.,11:3341-3351; and Qiu; et al., 1988, EMBO J., 7:1003-1011, which areincorporated by reference herein in their entirety, including anydrawings. Mutant versions of c-kit kinase are encompassed by the term“c-kit” or “c-kit kinase” and include those that fall into two classes:(1) having a single amino acid substitution at codon 816 of the humanc-kit kinase, or its equivalent position in other species (Ma et al.,1999, J. Invest Dermatol., 112:165-170), and (2) those which havemutations involving the putative juxtamembrane z-helix of the protein(Ma, et al., 1999, J. Biol. Chem., 274:13399-13402). Both of thesepublications are incorporated by reference herein in their entirety,including any drawings.

As used herein, “Bcr-Abl” is a fusion protein that results from thetranslocation of gene sequences from c-ABL protein tyrosine kinase onchromosome 9 into BCR sequences on chromosome 22 producing thePhiladelphia chromosome. A schematic representation of human Bcr, Abl,and Bcr-Abl can be seen in FIG. 1 of U.S. patent application Ser. No.10/193,651, filed on Jul. 9, 2002, the entire teachings of which areincorporated herein by reference. Depending on the breaking point in theBcr gene, Bcr-Abl fusion proteins can vary in size from 185-230 kDa butthey must contain at least the OLI domain from Bcr and the TK domainfrom Abl for transforming activity. The most common Bcr-Abl geneproducts found in humans are P230 Bcr-Abl, P210 Bcr-Abl, and P190Bcr-Abl. P210 Bcr-Abl is characteristic of CML and P190 Bcr-Abl ischaracteristic of ALL.

FLT3 kinase is a tyrosine kinase receptor involved in the regulation andstimulation of cellular proliferation (see Gilliland et al., Blood(2002), 100:1532-42, the entire teachings of which are incorporatedherein by reference). The FLT3 kinase has five immunoglobulin-likedomains in its extracellular region as well as an insert region of75-100 amino acids in the middle of its cytoplasmic domain. FLT3 kinaseis activated upon the binding of the FLT3 ligand, which causes receptordimerization. Dimerization of the FLT3 kinase by FLT3 ligand activatesthe intracellular kinase activity as well as a cascade of downstreamsubstrates including Stat5, Ras, phosphatidylinositol-3-kinase (PI3K),PLC□, Erk2, Akt, MAPK, SHC, SHP2, and SHIP (see Rosnet et al., ActaHaematol. (1996), 95:218; Hayakawa et al., Oncogene (2000), 19:624;Mizuki et al., Blood (2000), 96:3907; and Gilliand et al., Curr. Opin.Hematol. (2002), 9: 274-81, the entire teachings of each of thesereferences are incorporated herein by reference). Both membrane-boundand soluble FLT3 ligand bind, dimerize, and subsequently activate theFLT3 kinase.

Normal cells that express FLT3 kinase include immature hematopoieticcells, typically CD34+ cells, placenta, gonads, and brain (see Rosnet,et al., Blood (1993), 82:1110-19; Small et al., Proc. Natl. Acad. Sci.U.S.A. (1994), 91:459-63; and Rosnet et al., Leukemia (1996), 10:238-48,the entire teachings of each of these references are incorporated hereinby reference). However, efficient stimulation of proliferation via FLT3kinase typically requires other hematopoietic growth factors orinterleukins. FLT3 kinase also plays a critical role in immune functionthrough its regulation of dendritic cell proliferation anddifferentiation (see McKenna et al., Blood (2000), 95:3489-97, theentire teachings of which are incorporated herein by reference).

Numerous hematologic malignancies express FLT3 kinase, the mostprominent of which is AML (see Yokota et al., Leukemia (1997),11:1605-09, the entire teachings of which are incorporated herein byreference). Other FLT3 expressing malignancies include B-precursor cellacute lymphoblastic leukemias, myelodysplastic leukemias, T-cell acutelymphoblastic leukemias, and chronic myelogenous leukemias (see Rasko etal., Leukemia (1995), 9:2058-66, the entire teachings of which areincorporated herein by reference).

FLT3 kinase mutations associated with hematologic malignancies areactivating mutations. In other words, the FLT3 kinase is constitutivelyactivated without the need for binding and dimerization by FLT3 ligand,and therefore stimulates the cell to grow continuously. Two types ofactivating mutations have been identified: internal tandem duplications(ITDs) and point mutation in the activating loop of the kinase domain.As used herein, the term “FLT3 kinase” refers to both wild type FLT3kinase and mutant FLT3 kinases, such as FLT3 kinases that haveactivating mutations.

Compounds provided herein are useful in treating conditionscharacterized by inappropriate FLT3 activity such as proliferativedisorders. Inappropriate FLT3 activity includes, but is not limited to,enhanced FLT3 activity resulting from increased or de novo expression ofFLT3 in cells, increased FLT3 expression or activity, and FLT3 mutationsresulting in constitutive activation. The existence of inappropriate orabnormal FLT3 ligand and FLT3 levels or activity can be determined usingwell known methods in the art. For example, abnormally high FLT3 levelscan be determined using commercially available ELISA kits. FLT3 levelscan be determined using flow cytometric analysis, immunohistochemicalanalysis, and in situ hybridization techniques.

By “epidermal growth factor receptor” or “EGFR” as used herein is meant,any epidermal growth factor receptor (EGFR) protein, peptide, orpolypeptide having EGFR or EGFR family (e.g., HER1, HER2, HER3, and/orHER4) activity (such as encoded by EGFR Genbank Accession Nos. shown inTable I of U.S. patent application Ser. No. 10/923,354, filed on Aug.20, 2004, the entire teachings of which are incorporated herein byreference), or any other EGFR transcript derived from a EGFR gene and/orgenerated by EGFR translocation. The term “EGFR” is also meant toinclude other EGFR protein, peptide, or polypeptide derived from EGFRisoforms (e.g., HER1, HER2, HER3, and/or HER4), mutant EGFR genes,splice variants of EGFR genes, and EGFR gene polymorphisms.

As used herein, a “proliferative disorder” or a “hyperproliferativedisorder,” and other equivalent terms, means a disease or medicalcondition involving pathological growth of cells. Proliferativedisorders include cancer, smooth muscle cell proliferation, systemicsclerosis, cirrhosis of the liver, adult respiratory distress syndrome,idiopathic cardiomyopathy, lupus erythematosus, retinopathy, e.g.,diabetic retinopathy or other retinopathies, cardiac hyperplasia,reproductive system associated disorders such as benign prostatichyperplasia and ovarian cysts, pulmonary fibrosis, endometriosis,fibromatosis, harmatomas, lymphangiomatosis, sarcoidosis, desmoidtumors,

Smooth muscle cell proliferation includes hyperproliferation of cells inthe vasculature, for example, intimal smooth muscle cell hyperplasia,restenosis and vascular occlusion, particularly stenosis followingbiologically- or mechanically-mediated vascular injury, e.g., vascularinjury associated with angioplasty. Moreover, intimal smooth muscle cellhyperplasia can include hyperplasia in smooth muscle other than thevasculature, e.g., bile duct blockage, bronchial airways of the lung inpatients with asthma, in the kidneys of patients with renal interstitialfibrosis, and the like.

Non-cancerous proliferative disorders also include hyperproliferation ofcells in the skin such as psoriasis and its varied clinical forms,Reiter's syndrome, pityriasis rubra pilaris, and hyperproliferativevariants of disorders of keratinization (e.g., actinic keratosis, senilekeratosis), scleroderma, and the like.

In a preferred embodiment, the proliferative disorder is cancer. Cancersthat can be treated or prevented by the methods of the present inventioninclude, but are not limited to human sarcomas and carcinomas, e.g.,fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenicsarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma,lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor,leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer,breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma,basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceousgland carcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testiculartumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma,epithelial carcinoma, glioma, astrocytoma, medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma,retinoblastoma; leukemias, e.g., acute lymphocytic leukemia and acutemyelocytic leukemia (myeloblastic, promyelocytic, myelomonocytic,monocytic and erythroleukemia); chronic leukemia (chronic myelocytic(granulocytic) leukemia and chronic lymphocytic leukemia); andpolycythemia vera, lymphoma (Hodgkin's disease and non-Hodgkin'sdisease), multiple myeloma, Waldenstrobm's macroglobulinemia, and heavychain disease.

Other examples of leukemias include acute and/or chronic leukemias,e.g., lymphocytic leukemia (e.g., as exemplified by the p388 (murine)cell line), large granular lymphocytic leukemia, and lymphoblasticleukemia; T-cell leukemias, e.g., T-cell leukemia (e.g., as exemplifiedby the CEM, Jurkat, and HSB-2 (acute), YAC-1(murine) cell lines),T-lymphocytic leukemia, and T-lymphoblastic leukemia; B cell leukemia(e.g., as exemplified by the SB (acute) cell line), and B-lymphocyticleukemia; mixed cell leukemias, e.g., B and T cell leukemia and B and Tlymphocytic leukemia; myeloid leukemias, e.g., granulocytic leukemia,myelocytic leukemia (e.g., as exemplified by the HL-60 (promyelocyte)cell line), and myelogenous leukemia (e.g., as exemplified by the K562(chronic)cell line); neutrophilic leukemia; eosinophilic leukemia;monocytic leukemia (e.g., as exemplified by the THP-1 (acute) cellline); myelomonocytic leukemia; Naegeli-type myeloid leukemia; andnonlymphocytic leukemia. Other examples of leukemias are described inChapter 60 of The Chemotherapy Sourcebook, Michael C. Perry Ed.,Williams & Williams (1992) and Section 36 of Holland Frie CancerMedicine 5th Ed., Bast et al. Eds., B. C. Decker Inc. (2000). The entireteachings of the preceding references are incorporated herein byreference.

In one embodiment, the disclosed method is believed to be particularlyeffective in treating subject with non-solid tumors such as multiplemyeloma. In another embodiment, the disclosed method is believed to beparticularly effective against T-leukemia (e.g., as exemplified byJurkat and CEM cell lines); B-leukemia (e.g., as exemplified by the SBcell line); promyelocytes (e.g., as exemplified by the HL-60 cell line);uterine sarcoma (e.g., as exemplified by the MES-SA cell line);monocytic leukemia (e.g., as exemplified by the THP-1 (acute) cellline); and lymphoma (e.g., as exemplified by the U937 cell line).

In one embodiment, the disclosed method is believed to be particularlyeffective in treating subject with non-Hodgkin's lymphoma (NHL).Lymphomas are generally classified as either Hodgkin's disease (HD) ornon-Hodgkin's lymphomas (NHL). NHL differs from HD by the absence ofReed-Sternberg cells. The course of NHL is less predictable than HD andis more likely to spread to areas beyond the lymph nodes. NHL can befurther divided into B-cell NHL and T-cell NHL each of which can befurther categorized into a variety of different subtypes. For example,B-cell NHL includes Burkitt's lymphoma, follicular lymphoma, diffuselarge B-cell lymphoma, nodal marginal zone B-cell lymphoma, plasma cellneoplasms, small lymphocytic lymphoma/chronic lymphocytic leukemia,mantle cell lymphoma, extranodal marginal zone B-cell lymphoma, andlymphoplamacytic lymphoma/Waldenstrom macroglobulinemia. T-cell NHLinclude anaplastic large-cell lymphoma, precursor-T-cell lymphoblasticleukemia/lymphoma, unspecified peripheral T-cell lymphoma, acutelymphoblastic leukemia/lymphoma, angioimmunoblastic T-cell lymphoma, andmycosis fungoides.

Without wishing to be bound by any theory, it is believed that thecompounds of the invention are useful for treating NHLs, includingB-cell and T-cell NHLs, since Hsp90 is upregulated in many NHLs. Inparticular, in a survey of 412 cases of NHL in B-cell NHL, Hsp90 wasfound to be moderately to strongly over expressed in all cases ofBurkitt's lymphoma (5/5, 100%), and in a subset of follicular lymphoma(17/28, 61%), diffuse large B-cell lymphoma (27/46, 59%), nodal marginalzone B-cell lymphoma (6/16, 38%), plasma cell neoplasms (14/39, 36%),small lymphocytic lymphoma/chronic lymphocytic leukemia (3/9, 33%),mantle cell lymphoma (12/38, 32%), and lymphoplamacyticlymphoma/Waldenstrom macroglobulinemia (3/10, 30%). In addition, inT-cell NHL, Hsp90 was found to be moderately to strongly over expressedin a subset of anaplastic large-cell lymphoma (14/24, 58%),precursor-T-cell lymphoblastic leukemia/lymphoma (20/65, 31%),unspecified peripheral T-cell lymphoma (8/43, 23%), andangioimmunoblastic T-cell lymphoma (2/17, 12%). (See Valbuena, et al.,Modern Pathology (2005), 18:1343-1349, the entire teachings of which areincorporated herein by reference.)

Some of the disclosed methods can be particularly effective at treatingsubjects whose cancer has become “multi-drug resistant”. A cancer whichinitially responded to an anti-cancer drug becomes resistant to theanti-cancer drug when the anti-cancer drug is no longer effective intreating the subject with the cancer. For example, many tumors willinitially respond to treatment with an anti-cancer drug by decreasing insize or even going into remission, only to develop resistance to thedrug. Drug resistant tumors are characterized by a resumption of theirgrowth and/or reappearance after having seemingly gone into remission,despite the administration of increased dosages of the anti-cancer drug.Cancers that have developed resistance to two or more anti-cancer drugsare said to be “multi-drug resistant”. For example, it is common forcancers to become resistant to three or more anti-cancer agents, oftenfive or more anti-cancer agents and at times ten or more anti-canceragents.

As used herein, the term “c-kit associated cancer” refers to a cancerwhich has aberrant expression and/or activation of c-kit. c-Kitassociated cancers include leukemias, mast cell tumors, small cell lungcancer, testicular cancer, some cancers of the gastrointestinal tractand some central nervous system. In addition, c-kit has been implicatedin playing a role in carcinogenesis of the female genital tract (Inoue,et al., 1994, Cancer Res., 54(11):3049-3053), sarcomas ofneuroectodermal origin (Ricotti, et al., 1998, Blood, 91:2397-2405), andSchwann cell neoplasia associated with neurofibromatosis (Ryan, et al.,1994, J. Neuro. Res., 37:415-432).

In one embodiment, compounds of the invention are vascular targetingagents. In one aspect, compounds of the invention are effective forblocking, occluding, or otherwise disrupting blood flow in“neovasculature.” In one aspect, the invention provides a noveltreatment for diseases involving the growth of new blood vessels(“neovasculature”), including, but not limited to: cancer; infectiousdiseases; autoimmune disorders; benign tumors, e.g. hemangiomas,acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas;artheroscleric plaques; ocular angiogenic diseases, e.g., diabeticretinopathy, retinopathy of prematurity, macular degeneration, cornealgraft rejection, neovascular glaucoma, retrolental fibroplasia,rubeosis, retinoblastoma, persistent hyperplastic vitreous syndrome,choroidal neovascularization, uvietis and Pterygia (abnormal bloodvessel growth) of the eye; rheumatoid arthritis; psoriasis; warts;allergic dermatitis; blistering disease; Karposi sarcoma; delayed woundhealing; endometriosis; uterine bleeding; ovarian cysts; ovarianhyperstimulation; vasculogenesis; granulations; hypertrophic scars(keloids); nonunion fractures; scleroderma; trachoma; vascularadhesions; vascular malformations; DiGeorge syndrome; HHT; transplantarteriopathy; restinosis; obesity; myocardial angiogenesis; coronarycollaterals; cerebral collaterals; arteriovenous malformations; ischemiclimb angiogenesis; primary pulmonary hypertension; asthma; nasal polyps;inflammatory bowel disease; periodontal disease; ascites; peritonealadhesions; Osler-Webber Syndrome; plaque neovascularization;telangiectasia; hemophiliac joints; synovitis; osteomyelitis; osteophyteformation; angiofibroma; fibromuscular dysplasia; wound granulation;Crohn's disease; and atherosclerosis.

Vascular targeting can be demonstrated by any method known to thoseskilled in the art, such as the method described herein in Examples Eand F.

As used herein, the term “angiogenesis” refers to a fundamental processof generating new blood vessels in tissues or organs. Angiogenesis isinvolved with or associated with many diseases or conditions, including,but not limited to: cancer; ocular neovascular disease; age-relatedmacular degeneration; diabetic retinopathy, retinopathy of prematurity;corneal graft rejection; neovascular glaucoma; retrolental fibroplasias;epidemic keratoconjunctivitis; Vitamin A deficiency; contact lensoverwear; atopic keratitis; superior limbic keratitis; pterygiumkeratitis sicca; sjogrens; acne rosacea; warts; eczema; phylectenulosis;syphilis; Mycobacteria infections; lipid degeneration; chemical burns;bacterial ulcers; fungal ulcers; Herpes simplex infections; Herpeszoster infections; protozoan infections; Kaposi's sarcoma; Mooren'sulcer; Terrien's marginal degeneration; mariginal keratolysis;rheumatoid arthritis; systemic lupus; polyarteritis; trauma; Wegener'ssarcoidosis; scleritis; Stevens-Johnson disease; pemphigoid; radialkeratotomy; corneal graph rejection; diabetic retinopathy maculardegeneration; sickle cell anemia; sarcoid; syphilis; pseudoxanthomaelasticum; Paget's disease; vein occlusion; artery occlusion; carotidobstructive disease; chronic uveitis/vitritis; mycobacterial infections;Lyme's disease; systemic lupus erythematosis; retinopathy ofprematurity; Eales' disease; Behcet's disease; infections causing aretinitis or choroiditis; presumed ocular histoplasmosis; Best'sdisease; myopia; optic pits; Stargardt's disease; pars planitis; chronicretinal detachment; hyperviscosity syndromes; toxoplasmosis; trauma andpost-laser complications; diseases associated with rubeosis(neovasculariation of the angle); diseases caused by the abnormalproliferation of fibrovascular or fibrous tissue including all forms ofproliferative vitreoretinopathy; rheumatoid arthritis; osteoarthritis;ulcerative colitis; Crohn's disease; Bartonellosis; atherosclerosis;Osler-Weber-Rendu disease; hereditary hemorrhagic telangiectasia;pulmonary hemangiomatosis; pre-eclampsia; endometriosis; fibrosis of theliver and of the kidney; developmental abnormalities (organogenesis);skin disclolorations (e.g., hemangioma, nevus flammeus, or nevussimplex); wound healing; hypertrophic scars, i.e., keloids; woundgranulation; vascular adhesions; cat scratch disease (Rochele ninaliaquintosa); ulcers (Helicobacter pylori); keratoconjunctivitis;gingivitis; periodontal disease; epulis; hepatitis; tonsillitis;obesity; rhinitis; laryngitis; tracheitis; bronchitis; bronchiolitis;pneumonia; interstitial pulmonary fibrosis; neurodermitis; thyroiditis;thyroid enlargement; endometriosis; glomerulonephritis; gastritis;inflammatory bone and cartilage destruction; thromboembolic disease; andBuerger's disease.

The term “infection” is used herein in its broadest sense and refers toany infection e.g. a viral infection or one caused by a microorganism:bacterial infection, fungal infection, or parasitic infection (e.g.protozoal, amoebic, or helminth). Examples of such infections may befound in a number of well known texts such as “Medical Microbiology”(Greenwood, D., Slack, R., Peutherer, J., Churchill Livingstone Press,2002); “Mims' Pathogenesis of Infectious Disease” (Mims, C., Nash, A.,Stephen, J., Academic Press, 2000); “Fields” Virology. (Fields, B. N.,Knipe, D. M., Howley, P. M., Lippincott Williams and Wilkins, 2001); and“The Sanford Guide To Antimicrobial Therapy,” 26th Edition, J. P.Sanford et al. (Antimicrobial Therapy, Inc., 1996), all of which areincorporated by reference herein in their entirety.

“Bacterial infections” include, but are not limited to, infectionscaused by Gram Positive Bacteria including Bacillus cereus, Bacillusanthracis, Clostridium botulinum, Clostridium difficile, Clostridiumtetani, Clostridium perfringens, Corynebacteria diphtheriae,Enterococcus (Streptococcus D), Listeria monocytogenes, Pneumoccoccalinfections (Streptococcus pneumoniae), Staphylococcal infections andStreptococcal infections; Gram Negative Bacteria including Bacteroides,Bordetella pertussis, Brucella, Campylobacter infections,enterohaemorrhagic Escherichia coli (EHEC/E. coli 0157: H7)enteroinvasive Escherichia coli (EIEC); enterotoxigenic Escherichia coli(ETEC), Haemophilus influenzae, Helicobacter pylori, Klebsiellapneumoniae, Legionella spp., Moraxella catarrhalis, Neisseriagonnorrhoeae, Neisseria meningitidis, Proteus spp., Pseudomonasaeruginosa, Salmonella spp., Shigella spp., Vibrio cholera and Yersinia;acid fast bacteria including Mycobacterium tuberculosis, Mycobacteriumavium-intracellulare, Myobacterium johnei, Mycobacterium leprae,atypical bacteria, Chlamydia, Mycoplasma, Rickettsia, Spirochetes,Treponema pallidum, Borrelia recurrentis, Borrelia burgdorfii andLeptospira icterohemorrhagiae; or other miscellaneous bacteria,including Actinomyces and Nocardia.

The term “fungus” or “fungal” refers to a distinct group of eukaryotic,spore-forming organisms with absorptive nutrition and lackingchlorophyll. It includes mushrooms, molds, and yeasts.

“Fungal infections” include, but are not limited to, infections causedby Alternaria alternata, Aspergillus flavus, Aspergillus fumigatus,Aspergillus nidulans, Aspergillus niger, Aspergillus versicolor,Blastomyces dermatiditis, Candida albicans, Candida dubliensis, Candidakrusei, Candida parapsilosis, Candida tropicalis, Candida glabrata,Coccidioides immitis, Cryptococcus neoformans, Epidermophyton floccosum,Histoplasma capsulatum, Malassezia furfur, Microsporum canis, Mucorspp., Paracoccidioides brasiliensis, Penicillium marneffei, Pityrosporumovale, Pneumocystis carinii, Sporothrix schenkii, Trichophyton rubrum,Trichophyton interdigitale, Trichosporon beigelii, Rhodotorula spp.,Brettanomyces clausenii, Brettanomyces custerii, Brettanomycesanomalous, Brettanomyces naardenensis, Candida himilis, Candidaintermedia, Candida saki, Candida solani, Candida tropicalis, Candidaversatilis, Candida bechii, Candida famata, Candida lipolytica, Candidastellata, Candida vini, Debaromyces hansenii, Dekkera intermedia,Dekkera bruxellensis, Geotrichium sandidum, Hansenula fabiani,Hanseniaspora uvarum, Hansenula anomala, Hanseniaspora guillermondiiHanseniaspora vinae, Kluyveromyces lactis, Kloekera apiculata,Kluveromyces marxianus, Kluyveromyces fragilis, Metschikowiapulcherrima, Pichia guilliermodii, Pichia orientalis, Pichia fermentans,Pichia memranefaciens, Rhodotorula Saccharomyces bayanus, Saccharomycescerevisiae, Saccharomyces dairiensis Saccharomyces exigus, Saccharomycesuinsporus, Saccharomyces uvarum, Saccharomyces oleaginosus,Saccharomyces boulardii, Saccharomycodies ludwigii, Schizosaccharomycespombe, Torulaspora delbruekii, Torulopsis stellata, Zygoaccharomycesbailli and Zygosaccharomyces rouxii.

Drug resistance in fungi is characterized by the failure of anantifungal therapy to control a fungal infection. “Antifungalresistance” as used herein refers to both intrinsic or primary (presentbefore exposure to antifungal agents) and secondary or acquired(develops after exposure to antifungals). Hsp90 has been shown to play arole in the evolution of drug resistance in fungi. Cowen, L. et al.,Eukaryotic Cell, 2184-2188, 5(12), 2006; Cowen, L. et al., Science,309:2185-2189, 2005. It has been shown that the key mediator of Hsp90dependent azole resistance is calcineurin (a client protein of Hsp90).Calcineurin is required for tolerating the membrane stress exerted byazole drugs. Hsp90 keeps calcineurin stable and poised for activation.In addition, it has been shown that Hsp90 is required for the emergenceof drug resistance and continued drug resistance to azoles andechinocandins.

“Parasitic infections” include, but are not limited to, infectionscaused by Leishmania, Toxoplasma, Plasmodia, Theileria, Acanthamoeba,Anaplasma, Giardia, Trichomonas, Trypanosoma, Coccidia, and Babesia.

For example, parasitic infections include those caused by Trypanosomacruzi, Eimeria tenella, Plasmodium falciparum, Plasmodium vivax,Plasmodium ovale, Cryptosporidium parvum, Naegleria fowleri, Entamoebahistolytica, Balamuthia mandrillaris, Entameoba histolytica,Schistostoma mansoni, Plasmodium falciparum, P. vivax, P. ovale P.malariae, P. berghei, Leishmania donovani, L. infantum, L. chagasi, L.mexicana, L. amazonensis, L. venezuelensis, L. tropics, L. major, L.minor, L. aethiopica, L. Biana braziliensis, L. (V.) guyanensis, L. (V.)panamensis, L. (V.) peruviana, Trypanosoma brucei rhodesiense, T bruceigambiense, Giardia intestinalis, G. lambda, Toxoplasma gondii,Trichomonas vaginalis, Pneumocystis carinii, Acanthamoeba castellani A.culbertsoni, A. polyphaga, A. healyi, (A. astronyxis), A. hatchetti, A.rhysodes, and Trichinella spiralis.

As used herein, the term “viral infection” refers to any stage of aviral infection, including incubation phase, latent or dormant phase,acute phase, and development and maintenance of immunity towards avirus. Consequently, the term “treatment” is meant to include aspects ofgenerating or restoring immunity of the patient's immune system, as wellas aspects of suppressing or inhibiting viral replication.

Viral infections include, but are not limited to those caused byAdenovirus, Lassa fever virus (Arenavirus), Astrovirus, Hantavirus, RiftValley Fever virus (Phlebovirus), Calicivirus, Ebola virus, MarburgVirus, Japanese encephalitis virus, Dengue virus, Yellow fever virus,Hepatitis C virus, Hepatitis G virus, Hepatitis B virus, Hepatitis Dvirus, Herpes simplex virus 1, Herpes simplex virus 2), Cytomegalovirus,Epstein Barr virus, Varicella Zoster Virus, Human Herpesvirus 7, HumanHerpesvirus 8, Influenza virus, Parainfluenza virus, Rubella virus,Mumps virus, Morbillivirus, Measles virus, Respiratory Syncytial virus,Papillomaviruses, JC virus (Polyomavirus), BK virus (Polyomavirus),Parvovirus, Coxsackie virus (A and B), Hepatitis A virus, Polioviruses,Rhinoviruses, Reovirus, Rabies Virus (Lyssavirus), HumanImmunodeficiency virus 1 and 2, Human T-cell Leukemia virus.

Examples of viral infections include Adenovirus acute respiratorydisease, Lassa fever, Astrovirus enteritis, Hantavirus pulmonarysyndrome, Rift valley fever, Hepatitis E, diarrhoea, Ebola hemorrhagicfever, Marburg hemorrhagic fever, Japanese encephalitis, Dengue fever,Yellow fever, Hepatitis C, Hepatitis G, Hepatitis B, Hepatitis D, Coldsores, Genital sores, Cytomegalovirus infection, Mononucleosis, ChickenPox, Shingles, Human Herpesvirus infection 7, Kaposi Sarcoma, Influenza,Brochiolitis, German measles, Mumps, Measles (rubeola), Measles,Brochiolitis, Papillomas (Warts), cervical cancer, Progressivemultifocal leukoencephalopathy, Kidney disease, Erythema infectiosum,Viral myocarditis, meninigitis, entertitis, Hepatitis, Poliomyelitis,Cold, Diarrhoea, Rabies, AIDS and Leukemia.

DNA topoisomerases are enzymes present in all cells that catalyzetopological changes in DNA. Topoisomerase II (“topo II”) plays importantroles in DNA replication, chromosome segregation and the maintenance ofthe nuclear scaffold in eukaryotic cells. The enzyme acts by creatingbreaks in DNA, thereby allowing the DNA strands to unravel and separate.Due to the important roles of the enzyme in dividing cells, the enzymeis a highly attractive target for chemotherapeutic agents, especially inhuman cancers. The ability of compounds to inhibit topo II can bedetermined by any method known in the art such as in Example K.

The glucocorticoid receptor is a member of the steroid hormone nuclearreceptor family which includes glucocorticoid receptors (GR), androgenreceptors (AR), mineralocorticoid receptors (MR), estrogen receptors(ER), and progesterone receptors (PR). Glucocorticoid receptors bindglucocorticoids such as cortisol, corticosterone, and cortisone.

“Immunosuppression” refers to impairment of any component of the immunesystem resulting in decreased immune function. This impairment may bemeasured by any conventional means including whole blood assays oflymphocyte function, detection of lymphocyte proliferation andassessment of the expression of T cell surface antigens. The antisheepred blood cell (SRBC) primary (IgM) antibody response assay (usuallyreferred to as the plaque assay) is one specific method. This and othermethods are described in Luster, M. I., Portier, C., Pait, D. G., White,K. L., Jr., Gennings, C., Munson, A. E., and Rosenthal, G. J. (1992).“Risk Assessment in Immunotoxicology I: Sensitivity and Predictabilityof Immune Tests.” Fundam. Appl. Toxicol., 18, 200-210. Measuring theimmune response to a T-cell dependent immunogen is another particularlyuseful assay (Dean, J. H., House, R. V., and Luster, M. I. (2001).“Immunotoxicology: Effects of, and Responses to, Drugs and Chemicals.”In Principles and Methods of Toxicology: Fourth Edition (A. W. Hayes,Ed.), pp. 1415-1450, Taylor & Francis, Philadelphia, Pa.). In oneembodiment, a decrease in the expression of glucocorticoid receptors inPBMCs indicates impairment of immune function. A patient in need ofimmunosuppression is within the judgement of a physician, and caninclude patients with immune or inflammatory disorders. In oneembodiment, patients that have undergone or will be undergoing an organ,tissue, bone marrow, or stem cell transplantation are in need ofimmunosuppression to prevent inflammation and/or rejection of thetransplanted organ or tissue.

The compounds of this invention can be used to treat subjects withimmune disorders. As used herein, the term “immune disorder” and liketerms means a disease, disorder or condition caused by the immune systemof an animal, including autoimmune disorders. Immune disorders includethose diseases, disorders or conditions that have an immune componentand those that are substantially or entirely immune system-mediated.Autoimmune disorders are those wherein the animal's own immune systemmistakenly attacks itself, thereby targeting the cells, tissues, and/ororgans of the animal's own body. For example, the autoimmune reaction isdirected against the nervous system in multiple sclerosis and the gut inCrohn's disease. In other autoimmune disorders such as systemic lupuserythematosus (lupus), affected tissues and organs may vary amongindividuals with the same disease. One person with lupus may haveaffected skin and joints whereas another may have affected skin, kidney,and lungs. Ultimately, damage to certain tissues by the immune systemmay be permanent, as with destruction of insulin-producing cells of thepancreas in Type 1 diabetes mellitus. Specific autoimmune disorders thatmay be ameliorated using the compounds and methods of this inventioninclude without limitation, autoimmune disorders of the nervous system(e.g., multiple sclerosis, myasthenia gravis, autoimmune neuropathiessuch as Guillain-Barré, and autoimmune uveitis), autoimmune disorders ofthe blood (e.g., autoimmune hemolytic anemia, pernicious anemia, andautoimmune thrombocytopenia), autoimmune disorders of the blood vessels(e.g., temporal arteritis, anti-phospholipid syndrome, vasculitides suchas Wegener's granulomatosis, and Behcet's disease), autoimmune disordersof the skin (e.g., psoriasis, dermatitis herpetiformis, pemphigusvulgaris, and vitiligo), autoimmune disorders of the gastrointestinalsystem (e.g., Crohn's disease, ulcerative colitis, primary biliarycirrhosis, and autoimmune hepatitis), autoimmune disorders of theendocrine glands (e.g., Type 1 or immune-mediated diabetes mellitus,Grave's disease. Hashimoto's thyroiditis, autoimmune oophoritis andorchitis, and autoimmune disorder of the adrenal gland); and autoimmunedisorders of multiple organs (including connective tissue andmusculoskeletal system diseases) (e.g., rheumatoid arthritis, systemiclupus erythematosus, scleroderma, polymyositis, dermatomyositis,spondyloarthropathies such as ankylosing spondylitis, and Sjogren'ssyndrome). In addition, other immune system mediated diseases, such asgraft-versus-host disease and allergic disorders, are also included inthe definition of immune disorders herein. Because a number of immunedisorders are caused by inflammation, there is some overlap betweendisorders that are considered immune disorders and inflammatorydisorders. For the purpose of this invention, in the case of such anoverlapping disorder, it may be considered either an immune disorder oran inflammatory disorder. “Treatment of an immune disorder” hereinrefers to administering a compound represented by any of the formulasdisclosed herein to a subject, who has an immune disorder, a symptom ofsuch a disease or a predisposition towards such a disease, with thepurpose to cure, relieve, alter, affect, or prevent the autoimmunedisorder, the symptom of it, or the predisposition towards it.

As used herein, the term “allergic disorder” means a disease, conditionor disorder associated with an allergic response against normallyinnocuous substances. These substances may be found in the environment(such as indoor air pollutants and aeroallergens) or they may benon-environmental (such as those causing dermatological or foodallergies). Allergens can enter the body through a number of routes,including by inhalation, ingestion, contact with the skin or injection(including by insect sting). Many allergic disorders are linked toatopy, a predisposition to generate the allergic antibody IgE. BecauseIgE is able to sensitize mast cells anywhere in the body, atopicindividuals often express disease in more than one organ. For thepurpose of this invention, allergic disorders include anyhypersensitivity that occurs upon re-exposure to the sensitizingallergen, which in turn causes the release of inflammatory mediators.Allergic disorders include without limitation, allergic rhinitis (e.g.,hay fever), sinusitis, rhinosinusitis, chronic or recurrent otitismedia, drug reactions, insect sting reactions, latex reactions,conjunctivitis, urticaria, anaphylaxis and anaphylactoid reactions,atopic dermatitis, asthma and food allergies.

As used herein, the term “asthma” means a pulmonary disease, disorder orcondition characterized by reversible airway obstruction, airwayinflammation, and increased airway responsiveness to a variety ofstimuli.

Compounds represented by any of the formulas disclosed herein can beused to prevent or to treat subjects with inflammatory disorders. Asused herein, an “inflammatory disorder” means a disease, disorder orcondition characterized by inflammation of body tissue or having aninflammatory component. These include local inflammatory responses andsystemic inflammation. Examples of such inflammatory disorders include:transplant rejection, including skin graft rejection; chronicinflammatory disorders of the joints, including arthritis, rheumatoidarthritis, osteoarthritis and bone diseases associated with increasedbone resorption; inflammatory bowel diseases such as ileitis, ulcerativecolitis, Barrett's syndrome, and Crohn's disease; inflammatory lungdisorders such as asthma, adult respiratory distress syndrome, andchronic obstructive airway disease; inflammatory disorders of the eyeincluding corneal dystrophy; trachoma, onchocerciasis, uveitis,sympathetic ophthalmitis and endophthalmitis; chronic inflammatorydisorders of the gums, including gingivitis and periodontitis;tuberculosis; leprosy; inflammatory diseases of the kidney includinguremic complications, glomerulonephritis and nephrosis; inflammatorydisorders of the skin including sclerodermatitis, psoriasis and eczema;inflammatory diseases of the central nervous system, including chronicdemyelinating diseases of the nervous system, multiple sclerosis,AIDS-related neurodegeneration and Alzheimer's disease, infectiousmeningitis, encephalomyelitis, Parkinson's disease, Huntington'sdisease, amyotrophic lateral sclerosis and viral or autoimmuneencephalitis; autoimmune disorders, immune-complex vasculitis, systemiclupus and erythematodes; systemic lupus erythematosus (SLE); andinflammatory diseases of the heart such as cardiomyopathy, ischemicheart disease hypercholesterolemia, atherosclerosis; as well as variousother diseases with significant inflammatory components, includingpreeclampsia; chronic liver failure, brain and spinal cord trauma. Theremay also be a systemic inflammation of the body, exemplified bygam-positive or gram negative shock, hemorrhagic or anaphylactic shock,or shock induced by cancer chemotherapy in response to pro-inflammatorycytokines, e.g., shock associated with pro-inflammatory cytokines. Suchshock can be induced, e.g., by a chemotherapeutic agent used in cancerchemotherapy. “Treatment of an inflammatory disorder” herein refers toadministering a compound or a composition of the invention to a subject,who has an inflammatory disorder, a symptom of such a disorder or apredisposition towards such a disorder, with the purpose to cure,relieve, alter, affect, or prevent the inflammatory disorder, thesymptom of it, or the predisposition towards it.

As used herein, the term “pharmaceutically acceptable salt,” is a saltformed from, for example, an acid and a basic group of one of thecompounds of formula (I)-(VIII), or Table 1. Illustrative salts include,but are not limited, to sulfate, citrate, acetate, oxalate, chloride,bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate,isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate,tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,besylate, gentisinate, fumarate, gluconate, glucaronate, saccharate,formate, benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate, and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. The term“pharmaceutically acceptable salt” also refers to a salt prepared from acompound of formula (I)-(VIII), or Table 1 having an acidic functionalgroup, such as a carboxylic acid functional group, and apharmaceutically acceptable inorganic or organic base. Suitable basesinclude, but are not limited to, hydroxides of alkali metals such assodium, potassium, and lithium; hydroxides of alkaline earth metal suchas calcium and magnesium; hydroxides of other metals, such as aluminumand zinc; ammonia, and organic amines, such as unsubstituted orhydroxy-substituted mono-, di-, or trialkylamines; dicyclohexylamine;tributyl amine; pyridine; N-methyl, N-ethylamine; diethylamine;triethylamine; mono-, bis-, or tris-(2-hydroxy-lower alkyl amines), suchas mono-, bis-, or tris-(2-hydroxyethyl)amine,2-hydroxy-tert-butylamine, or tris-(hydroxymethyl)methylamine,N,N,-di-lower alkyl-N-(hydroxy lower alkyl)-amines, such asN,N-dimethyl-N-(2-hydroxyethyl)amine, or tri-(2-hydroxyethyl)amine;N-methyl-D-glucamine; and amino acids such as arginine, lysine, and thelike. The term “pharmaceutically acceptable salt” also refers to a saltprepared from a compound of formula (I)-(VIII), or Table 1 having abasic functional group, such as an amine functional group, and apharmaceutically acceptable inorganic or organic acid. Suitable acidsinclude, but are not limited to, hydrogen sulfate, citric acid, aceticacid, oxalic acid, hydrochloric acid (HCl), hydrogen bromide (HBr),hydrogen iodide (HI), nitric acid, hydrogen bisulfide, phosphoric acid,lactic acid, salicylic acid, tartaric acid, bitartratic acid, ascorbicacid, succinic acid, maleic acid, besylic acid, fumaric acid, gluconicacid, glucaronic acid, formic acid, benzoic acid, glutamic acid,methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, andp-toluenesulfonic acid.

As used herein, the term “pharmaceutically acceptable solvate,” is asolvate formed from the association of one or more pharmaceuticallyacceptable solvent molecules to one of the compounds of formula(I)-(VIII), or Table 1. The term solvate includes hydrates (e.g.,hemihydrate, monohydrate, dihydrate, trihydrate, tetrahydrate, and thelike).

A pharmaceutically acceptable carrier may contain inert ingredientswhich do not unduly inhibit the biological activity of the compounds.The pharmaceutically acceptable carriers should be biocompatible, i.e.,non-toxic, non-inflammatory, non-immunogenic and d devoid of otherundesired reactions upon the administration to a subject. Standardpharmaceutical formulation techniques can be employed, such as thosedescribed in Remington's Pharmaceutical Sciences, ibid. Suitablepharmaceutical carriers for parenteral administration include, forexample, sterile water, physiological saline, bacteriostatic saline(saline containing about 0.9% mg/ml benzyl alcohol), phosphate-bufferedsaline, Hank's solution, Ringer's-lactate and the like. Methods forencapsulating compositions (such as in a coating of hard gelatin orcyclodextran) are known in the art (Baker, et al., “Controlled Releaseof Biological Active Agents”, John Wiley and Sons, 1986).

As used herein, the term “effective amount” refers to an amount of acompound of this invention which is sufficient to reduce or amelioratethe severity, duration, progression, or onset of a disease or disorder,e.g. a proliferative disorder, prevent the advancement of a a disease ordisorder, e.g. a proliferative disorder, cause the regression of a adisease or disorder, e.g. a proliferative disorder, prevent therecurrence, development, onset or progression of a symptom associatedwith a a disease or disorder, e.g. a proliferative disorder, or enhanceor improve the prophylactic or therapeutic effect(s) of another therapy.The precise amount of compound administered to a subject will depend onthe mode of administration, the type and severity of the disease orcondition and on the characteristics of the subject, such as generalhealth, age, sex, body weight and tolerance to drugs. It will alsodepend on the degree, severity and type of cell proliferation, and themode of administration. The skilled artisan will be able to determineappropriate dosages depending on these and other factors. Whenco-administered with other agents, e.g., when co-administered with ananti-cancer agent, an “effective amount” of the second agent will dependon the type of drug used. Suitable dosages are known for approved agentsand can be adjusted by the skilled artisan according to the condition ofthe subject, the type of condition(s) being treated and the amount of acompound of the invention being used. In cases where no amount isexpressly noted, an effective amount should be assumed.

Non-limiting examples of an effective amount of a compound of theinvention are provided herein below. In a specific embodiment, theinvention provides a method of preventing, treating, managing, orameliorating a proliferative disorder or one or more symptoms thereof,said methods comprising administering to a subject in need thereof adose of at least 150 μg/kg, preferably at least 250 μg/kg, at least 500μg/kg, at least 1 mg/kg, at least 5 mg/kg, at least 10 mg/kg, at least25 mg/kg, at least 50 mg/kg, at least 75 mg/kg, at least 100 mg/kg, atleast 125 mg/kg, at least 150 mg/kg, or at least 200 mg/kg or more ofone or more compounds of the invention once every day, preferably, onceevery 2 days, once every 3 days, once every 4 days, once every 5 days,once every 6 days, once every 7 days, once every 8 days, once every 10days, once every two weeks, once every three weeks, or once a month.

The dosages of a chemotherapeutic agents other than compounds of theinvention, which have been or are currently being used to prevent,treat, manage, or ameliorate a proliferative disorder, or one or moresymptoms thereof, can be used in the combination therapies of theinvention. Preferably, dosages lower than those which have been or arecurrently being used to prevent, treat, manage, or ameliorate aproliferative disorder, or one or more symptoms thereof, are used in thecombination therapies of the invention. The recommended dosages ofagents currently used for the prevention, treatment, management, oramelioration of a proliferative disorder, or one or more symptomsthereof, can obtained from any reference in the art including, but notlimited to, Hardman et al., eds., 1996, Goodman & Gilman's ThePharmacological Basis Of Basis Of Therapeutics 9^(th) Ed, Mc-Graw-Hill,New York; Physician's Desk Reference (PDR) 57^(th) Ed., 2003, MedicalEconomics Co., Inc., Montvale, N.J., which are incorporated herein byreference in its entirety.

As used herein, the terms “treat”, “treatment” and “treating” refer tothe reduction or amelioration of the progression, severity and/orduration of a a disease or disorder, e.g. a proliferative disorder, orthe amelioration of one or more symptoms (preferably, one or morediscernible symptoms) of a a disease or disorder, a proliferativedisorder resulting from the administration of one or more therapies(e.g., one or more therapeutic agents such as a compound of theinvention). In specific embodiments, the terms “treat”, “treatment” and“treating” refer to the amelioration of at least one measurable physicalparameter of a a disease or disorder, e.g. a proliferative disorder,such as growth of a tumor, not necessarily discernible by the patient.In other embodiments the terms “treat”, “treatment” and “treating” referto the inhibition of the progression of a a disease or disorder, e.g. aproliferative disorder, either physically by, e.g., stabilization of adiscernible symptom, physiologically by, e.g., stabilization of aphysical parameter, or both. In other embodiments the terms “treat”,“treatment” and “treating” refer to the reduction or stabilization oftumor size or cancerous cell count.

As used herein, the terms “prevent”, “prevention” and “preventing” referto the reduction in the risk of acquiring or developing a given adisease or disorder, e.g. a proliferative disorder, or the reduction orinhibition of the recurrence or a a disease or disorder, e.g. aproliferative disorder. In one embodiment, a compound of the inventionis administered as a preventative measure to a patient, preferably ahuman, having a genetic predisposition to any of the disorders describedherein.

As used herein, the terms “therapeutic agent” and “therapeutic agents”refer to any agent(s) which can be used in the treatment, management, oramelioration of a a disease or disorder, e.g. a proliferative disorderor one or more symptoms thereof. In certain embodiments, the term“therapeutic agent” refers to a compound of the invention. In certainother embodiments, the term “therapeutic agent” refers does not refer toa compound of the invention. Preferably, a therapeutic agent is an agentwhich is known to be useful for, or has been or is currently being usedfor the treatment, management, prevention, or amelioration a a diseaseor disorder, e.g. a proliferative disorder or one or more symptomsthereof.

As used herein, the term “synergistic” refers to a combination of acompound of the invention and another therapy (e.g., a prophylactic ortherapeutic agent), which is more effective than the additive effects ofthe therapies. A synergistic effect of a combination of therapies (e.g.,a combination of prophylactic or therapeutic agents) permits the use oflower dosages of one or more of the therapies and/or less frequentadministration of said therapies to a subject with a disease ordisorder, e.g. a proliferative disorder. The ability to utilize lowerdosages of a therapy (e.g., a prophylactic or therapeutic agent) and/orto administer said therapy less frequently reduces the toxicityassociated with the administration of said therapy to a subject withoutreducing the efficacy of said therapy in the prevention, management ortreatment of a disease or disorder, e.g. a proliferative disorder. Inaddition, a synergistic effect can result in improved efficacy of agentsin the prevention, management or treatment of a disease or disorder,e.g. a proliferative disorder. Finally, a synergistic effect of acombination of therapies (e.g., a combination of prophylactic ortherapeutic agents) may avoid or reduce adverse or unwanted side effectsassociated with the use of either therapy alone.

As used herein, the phrase “side effects” encompasses unwanted andadverse effects of a therapy (e.g., a prophylactic or therapeuticagent). Side effects are always unwanted, but unwanted effects are notnecessarily adverse. An adverse effect from a therapy (e.g.,prophylactic or therapeutic agent) might be harmful or uncomfortable orrisky. Side effects include, but are not limited to fever, chills,lethargy, gastrointestinal toxicities (including gastric and intestinalulcerations and erosions), nausea, vomiting, neurotoxicities,nephrotoxicities, renal toxicities (including such conditions aspapillary necrosis and chronic interstitial nephritis), hepatictoxicities (including elevated serum liver enzyme levels),myelotoxicities (including leukopenia, myelosuppression,thrombocytopenia and anemia), dry mouth, metallic taste, prolongation ofgestation, weakness, somnolence, pain (including muscle pain, bone painand headache), hair loss, asthenia, dizziness, extra-pyramidal symptoms,akathisia, cardiovascular disturbances and sexual dysfunction.

As used herein, the term “in combination” refers to the use of more thanone therapies (e.g., one or more prophylactic and/or therapeuticagents). The use of the term “in combination” does not restrict theorder in which therapies (e.g., prophylactic and/or therapeutic agents)are administered to a subject with a proliferative disorder. A firsttherapy (e.g., a prophylactic or therapeutic agent such as a compound ofthe invention) can be administered prior to (e.g., 5 minutes, 15minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks,4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantlywith, or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6weeks, 8 weeks, or 12 weeks after) the administration of a secondtherapy (e.g., a prophylactic or therapeutic agent such as ananti-cancer agent) to a subject with a proliferative disorder, such ascancer.

As used herein, the terms “therapies” and “therapy” can refer to anyprotocol(s), method(s), and/or agent(s) that can be used in theprevention, treatment, management, or amelioration of a a disease ordisorder, e.g. a proliferative disorder or one or more symptoms thereof.

A used herein, a “protocol” includes dosing schedules and dosingregimens. The protocols herein are methods of use and includeprophylactic and therapeutic protocols.

As used herein, the terms “manage,” “managing,” and “management” referto the beneficial effects that a subject derives from a therapy (e.g., aprophylactic or therapeutic agent), which does not result in a cure ofthe disease. In certain embodiments, a subject is administered one ormore therapies (e.g., one or more prophylactic or therapeutic agents) to“manage” a disease so as to prevent the progression or worsening of thedisease.

As used herein, a composition that “substantially” comprises a compoundmeans that the composition contains more than about 80% by weight, morepreferably more than about 90% by weight, even more preferably more thanabout 95% by weight, and most preferably more than about 97% by weightof the compound.

As used herein, a reaction that is “substantially complete” means thatthe reaction contains more than about 80% by weight of the desiredproduct, more preferably more than about 90% by weight of the desiredproduct, even more preferably more than about 95% by weight of thedesired product, and most preferably more than about 97% by weight ofthe desired product.

As used herein, a racemic mixture means about 50% of one enantiomer andabout 50% of is corresponding enantiomer relative to a chiral center inthe molecule. The invention encompasses all enantiomerically-pure,enantiomerically-enriched, diastereomerically pure, diastereomericallyenriched, and racemic mixtures of the compounds of the invention.

Enantiomeric and diastereomeric mixtures can be resolved into theircomponent enantiomers or diastereomers by well known methods, such aschiral-phase gas chromatography, chiral-phase high performance liquidchromatography, crystallizing the compound as a chiral salt complex, orcrystallizing the compound in a chiral solvent. Enantiomers anddiastereomers can also be obtained from diastereomerically- orenantiomerically-pure intermediates, reagents, and catalysts by wellknown asymmetric synthetic methods.

The compounds of the invention are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

When administered to a patient, e.g., to a non-human animal forveterinary use or for improvement of livestock, or to a human forclinical use, the compounds of the invention are administered inisolated form or as the isolated form in a pharmaceutical composition.As used herein, “isolated” means that the compounds of the invention areseparated from other components of either (a) a natural source, such asa plant or cell, preferably bacterial culture, or (b) a syntheticorganic chemical reaction mixture. Preferably, the compounds of theinvention are purified via conventional techniques. As used herein,“purified” means that when isolated, the isolate contains at least 95%,preferably at least 98%, of a compound of the invention by weight of theisolate either as a mixture of stereoisomers or as a diastereomeric orenantiomeric pure isolate.

As used herein, a composition that is “substantially free” of a compoundmeans that the composition contains less than about 20% by weight, morepreferably less than about 10% by weight, even more preferably less thanabout 5% by weight, and most preferably less than about 3% by weight ofthe compound.

Only those choices and combinations of substituents that result in astable structure are contemplated. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

The invention can be understood more fully by reference to the followingdetailed description and illustrative examples, which are intended toexemplify non-limiting embodiments of the invention.

B. The Compounds of the Invention

The present invention encompasses compounds having formulas (I)-(VIII),and those set forth in Table 1 and tautomers, pharmaceuticallyacceptable salts, solvates, clathrates, hydrates, polymorphs andprodrugs thereof.

Compounds of formulas (I)-(VIII), inhibit the activity of Hsp90 and areparticularly useful for treating or preventing proliferative disorders,such as cancer. In addition, compounds of formula (I)-(VIII), areparticularly useful in treating cancer when given in combination withanother anti-cancer agent.

In one embodiment, the invention provides compounds of formula (I) asset forth below:

wherein:

-   -   R₁, R₂ and R₃ are independently —OH, —SH, —NR₇H, —OR₂₆, —SR₂₆,        —O(CH₂)_(m)OH, —O(CH₂)_(m)SH, —O(CH₂)_(m)NR₇H, —S(CH₂)_(m)OH,        —S(CH₂)_(m)SH, —S(CH₂)_(m)NR₇H, —OC(O)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁,        —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇, —OC(O)OR₇,        —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇, —NR₇CH₂C(O)R₇,        —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇, —OCH₂C(O)NR₁₀R₁₁,        —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,        —SS(O)_(p)R₇, —S(O)_(p)OR₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁,        —SS(O)_(p)NR₁₀R₁₁, —NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇,        —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇, —OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇,        —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇, —OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁,        —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇, —NR₇C(NR₈)R₇,        —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,        —SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —OP(O)(OR₇)₂, or        —SP(O)(OR₇)₂, provided that at least one of R₁, R₂ and R₃ is        —OP(O)(OH)₂;    -   R₅ is —X₂₀R₅₀, an optionally substituted alkyl, an optionally        substituted alkenyl, an optionally substituted alkynyl, an        optionally substituted cycloalkyl, an optionally substituted        cycloalkenyl, an optionally substituted heterocyclyl, an        optionally substituted aryl, an optionally substituted        heteroaryl, an optionally substituted aralkyl, or an optionally        substituted heteraralkyl;    -   R₇ and R₈, for each occurrence, is independently, —H, an        optionally substituted alkyl, an optionally substituted alkenyl,        an optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;    -   R₁₀ and R₁₁, for each occurrence, is independently —H, an        optionally substituted alkyl, an optionally substituted alkenyl,        an optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;        or R₁₀ and R₁₁, taken together with the nitrogen to which they        are attached, form an optionally substituted heterocyclyl or an        optionally substituted heteroaryl;    -   R₂₆ is a lower alkyl;    -   R₅₀ is an optionally substituted aryl or an optionally        substituted heteroaryl;    -   X₂₀ is a C1-C4 alkyl, NR₇, C(O), C(S), C(NR₈), or S(O)_(p);    -   Z is a substituent;    -   p, for each occurrence, is independently, 1 or 2;    -   m for each occurrence, is independently 1, 2, 3, or 4; and    -   n is 0, 1, 2, or 3;    -   or a tautomer, pharmaceutically acceptable salt, solvate,        clathrate or a prodrug thereof.

In one embodiment, of the compounds represented by formula (I), thecompound is not3-hydroxy-4-(5-mercapto-4-(naphthalen-1-yl)-4H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate.

In one embodiment, the invention provides compounds of formula (II) asset forth below:

-   -   or a tautomer, pharmaceutically acceptable salt, solvate,        clathrate or a prodrug thereof, wherein R₆ is an optionally        substituted alkyl, an optionally substituted alkenyl, an        optionally substituted alkynyl, cyano, halo, nitro, an        optionally substituted cycloalkyl, haloalkyl, an optionally        substituted heterocyclyl, an optionally substituted aryl, an        optionally substituted heteroaryl, an optionally substituted        aralkyl, an optionally substituted heteroaralkyl, —OR₇, —SR₇,        —OC(O)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁, —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇,        —SC(O)R₇, —NR₇C(O)R₇, —OC(O)OR₇, —SC(O)OR₇, —NR₇C(O)OR₇,        —OCH₂C(O)R₇, —SCH₂C(O)R₇, —NR₇CH₂C(O)R₇, —OCH₂C(O)OR₇,        —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇, —OCH₂C(O)NR₁₀R₁₁,        —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,        —SS(O)_(p)R₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁,        —SS(O)_(p)NR₁₀R₁₁, —NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇,        —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇, —OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇,        —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇, —OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁,        —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇, —NR₇C(NR₈)R₇,        —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,        —SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —C(O)R₇, —C(O)OR₇,        —C(O)NR₁₀R₁₁, —C(O)SR₇, —C(S)R₇, —C(S)OR₇, —C(S)NR₁₀R₁₁,        —C(S)SR₇, —C(NR₈)OR₇, —C(NR₈)R₇, —C(NR₈)NR₁₀R₁₁, —C(NR₈)SR₇,        —S(O)_(p)OR₇, —S(O)_(p)NR₁₀OR₁₁, or —S(O)_(p)R₇; and R₁, R₂, R₃,        and R₅ are defined as for formula (I).

In one embodiment, the invention provides compounds of formula (III) asset forth below:

or a tautomer, pharmaceutically acceptable salt, solvate, clathrate or aprodrug thereof, wherein R₁, R₃, and R₅ are defined as for formula (I)and R₆ is defined as for formula (II).

In one embodiment, the invention provides compounds of formula (IV) asset forth below:

or a tautomer, pharmaceutically acceptable salt, solvate, clathrate or aprodrug thereof, wherein R₅ is defined as for formula (I) and R₆ isdefined as for formula (II).

In one embodiment, the invention provides compounds of formula (V) asset forth below:

or a tautomer, pharmaceutically acceptable salt, solvate, clathrate or aprodrug thereof,wherein

-   -   X₄₁ is O, S, or NR₄₂;    -   X₄₂ is CR₄₄ or N;    -   Y₄₀ is N or CR₄₃;    -   Y₄₁ is N or CR₄₅;    -   Y₄₂, for each occurrence, is independently N, C or CR₄₆;    -   R₄₁ is an optionally substituted alkyl, an optionally        substituted alkenyl, an optionally substituted alkynyl, cyano,        halo, nitro, an optionally substituted cycloalkyl, haloalkyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, an optionally substituted heteroaralkyl,        —OR₇, —SR₇, —NR₁₀R₁₁, —OC(O)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁,        —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇, —OC(O)OR₇,        —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇, —NR₇CH₂C(O)R₇,        —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇, —OCH₂C(O)NR₁₀R₁₁,        —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,        —SS(O)_(p)R₇, —NR₇S(O)_(p)OR₇, —OS(O)_(p)NR₁₀R₁₁,        —SS(O)_(p)NR₁₀OR₁₁, —NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇,        —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇, —OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇,        —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇, —OC(S)NR₁₀R₁₁,        —SC(S)NR₁₀OR₁₁, —NR₇C(S)NR₁₀OR₁₁—OC(NR₈)R₇, —SC(NR₈)R₇,        —NR₇C(NR₈)R₇, —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇,        —OC(NR₈)NR₁₀R₁₁, —SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —C(O)R₇,        —C(O)OR₇, —C(O)NR₁₀R₁₁, —C(O)SR₇, —C(S)R₇, —C(S)OR₇,        —C(S)NR₁₀OR₁₁, —C(S)SR₇, —C(NR₈)OR₇, —C(NR₈)R₇, —C(NR₈)NR₁₀OR₁₁,        —C(NR₈)SR₇, —S(O)_(p)OR₇, —S(O)_(p)NR₁₀R₁₁, or —S(O)_(p)R₇;    -   R₄₂ is —H, an optionally substituted alkyl, an optionally        substituted alkenyl, an optionally substituted alkynyl, an        optionally substituted cycloalkyl, an optionally substituted        cycloalkenyl, an optionally substituted heterocyclyl, an        optionally substituted aryl, an optionally substituted        heteroaryl, an optionally substituted aralkyl, an optionally        substituted heteraralkyl, hydroxyalkyl, alkoxyalkyl, a        haloalkyl, a heteroalkyl, —C(O)R₇, —(CH₂)_(m)C(O)OR₇, —C(O)OR₇,        —OC(O)R₇, —C(O)NR₁₀R₁₁, —S(O)_(p)R₇, —S(O)_(p)OR₇, or        —S(O)_(p)NR₁₀R₁₁;    -   R₄₃ and R₄₄ are, independently, —H, —OH, an optionally        substituted alkyl, an optionally substituted alkenyl, an        optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, an optionally substituted heteraralkyl,        hydroxyalkyl, alkoxyalkyl, halo, cyano, nitro, guanadino, a        haloalkyl, a heteroalkyl, —C(O)R₇, —C(O)OR₇, —OC(O)R₇,        —C(O)NR₁₀R₁₁, —NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇,        —S(O)_(p)OR₇, —NR₈S(O)_(p)R₇, —S(O)_(p)NR₁₀OR₁₁, or R₄₃ and R₄₄        taken together with the carbon atoms to which they are attached        form an optionally substituted cycloalkenyl, an optionally        substituted aryl, an optionally substituted heterocyclyl, or an        optionally substituted heteroaryl;    -   R₄₅ is —H, —OH, —SH, —NR₇H, —OR₂₆, —SR₂₆, —NHR₂₆, —O(CH₂)_(m)OH,        —O(CH₂)_(m), —SH, —O(CH₂)_(m)NR₇H, —S(CH₂)_(m)OH, —S(CH₂)_(m)SH,        —S(CH₂)_(m)NR₇H, —OC(O)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁, —NR₇C(O)NR₁₀R₁₁,        —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇, —OC(O)OR₇, —SC(O)OR₇,        —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇, —NR₇CH₂C(O)R₇,        —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇, —OCH₂C(O)NR₁₀OR₁₁,        —SCH₂C(O)NR₁₀OR₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,        —SS(O)_(p)R₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁,        —SS(O)_(p)NR₁₀R₁₁, —NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇,        —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇, —OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇,        —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇, —OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁,        —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇, —NR₇C(NR₈)R₇,        —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,        —SC(NR₈)NR₁₀R₁₁, or —NR₇C(NR₈)NR₁₀R₁₁; and    -   R₄₆, for each occurrence, is independently selected from the        group consisting of H, an optionally substituted alkyl, an        optionally substituted alkenyl, an optionally substituted        alkynyl, an optionally substituted cycloalkyl, an optionally        substituted cycloalkenyl, an optionally substituted        heterocyclyl, an optionally substituted aryl, an optionally        substituted heteroaryl, an optionally substituted aralkyl, an        optionally substituted heteraralkyl, halo, cyano, nitro,        guanadino, a haloalkyl, a heteroalkyl, —NR₁₀OR₁₁, —OR₇, —C(O)R₇,        —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁, —NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇,        —OS(O)_(p)R₇, —S(O)_(p)OR₇, —NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁.

In one embodiment, the invention provides compounds of formula (VI) asset forth below:

or a tautomer, pharmaceutically acceptable salt, solvate, clathrate or aprodrug thereof, wherein R₄₁, R₄₂, R₄₃, R₄₅, and X₄₂ are defined as forformula (V).

In one embodiment; the invention provides compounds of formula (VII) asset forth below:

or a tautomer, pharmaceutically acceptable salt, solvate, clathrate or aprodrug thereof, wherein

-   -   X₄₅ is CR₅₄ or N;    -   R₅₂ is selected from the group consisting of —H, methyl, ethyl,        n-propyl, isopropyl, n-butyl, n-pentyl, n-hexyl, —(CH₂)₂OCH₃,        —CH₂C(O)OH, and —C(O)N(CH₃)₂;    -   R₅₃ and R₅₄ are each, independently, —H, methyl, ethyl, or        isopropyl; or R₅₃ and R₅₄ taken together with the carbon atoms        to which they are attached form a phenyl, cyclohexenyl, or        cyclooctenyl ring;    -   R₅₅ is selected from the group consisting of —H, —OH, —OCH₃, and        —OCH₂CH₃; and    -   R₅₆ is selected from the group consisting of methyl, ethyl,        isopropyl, and cyclopropyl.

In one embodiment, the invention provides compounds of formula (VIII) asset forth below:

or a tautomer, pharmaceutically acceptable salt, solvate, clathrate or aprodrug thereof, wherein R₄₁ and R₄₂ are defined as for formula (V).

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is represented by the following formula:

-   -   wherein:    -   R₉, for each occurrence, is independently a substituent selected        from the group consisting of an optionally substituted alkyl, an        optionally substituted alkenyl, an optionally substituted        alkynyl, an optionally substituted cycloalkyl, an optionally        substituted cycloalkenyl, an optionally substituted        heterocyclyl, an optionally substituted aryl, an optionally        substituted heteroaryl, an optionally substituted aralkyl, an        optionally substituted heteraralkyl, hydroxyalkyl, alkoxyalkyl,        halo, cyano, nitro, guanadino, a haloalkyl, a heteroalkyl,        —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀OR₁₁,        —NR₈C(O)R₇, —SR_(S), —S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,        —NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —OP(O)(OR₇)₂,        or —SP(O)(OR₇)₂; or two R₉ groups taken together with the carbon        atoms to which they are attached form a fused ring; and    -   q is zero or an integer from 1 to 7.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is represented by the following formula:

-   -   wherein:    -   q is zero or an integer from 1 to 5; and    -   u is zero or an integer from 1 to 5.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is represented by the following formula:

-   -   wherein:    -   R₃₃ is a halo, lower alkyl, a lower alkoxy, a lower haloalkyl, a        lower haloalkoxy, and lower alkyl sulfanyl;    -   R₃₄ is H, a lower alkyl, or a lower alkylcarbonyl; and    -   Ring B and Ring C are optionally substituted with one or more        substituents.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is selected from the group consisting of:

-   -   wherein:    -   X₆, for each occurrence, is independently CH, CR₉, N,N(O),        N⁺(R₁₇), provided that at least three X₆ groups are        independently selected from CH and CR₉;    -   X₇, for each occurrence, is independently CH, CR₉, N,N(O),        N⁺(R₁₇), provided that at least three X₇ groups are        independently selected from CH and CR₉;    -   X₈, for each occurrence, is independently CH₂, CHR₉, C(R₉)₂, S,        S(O)_(p), NR₇, or NR₁₇;    -   X₉, for each occurrence, is independently N or CH;    -   X₁₀, for each occurrence, is independently CH, CR₉, N,N(O),        N⁺(R₁₇), provided that at least one X₁₀ is selected from CH and        CR₉;    -   R₉, for each occurrence, is independently a substituent selected        from the group consisting of an optionally substituted alkyl, an        optionally substituted alkenyl, an optionally substituted        alkynyl, an optionally substituted cycloalkyl, an optionally        substituted cycloalkenyl, an optionally substituted        heterocyclyl, an optionally substituted aryl, an optionally        substituted heteroaryl, an optionally substituted aralkyl, an        optionally substituted heteraralkyl, hydroxyalkyl, alkoxyalkyl,        halo, cyano, nitro, guanadino, a haloalkyl, a heteroalkyl,        —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁,        —NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,        —NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —OP(O)(OR₇)₂,        or —SP(O)(OR₇)₂, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂;        or two R₉ groups taken together with the carbon atoms to which        they are attached form a fused ring; and    -   R₁₇, for each occurrence, is independently —H, an alkyl, an        aralkyl, —C(O)R₇, —C(O)OR₇, or —C(O)NR₁₀R₁₁.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is an optionally substituted indolyl, an optionallysubstituted benzoimidazolyl, an optionally substituted indazolyl, anoptionally substituted 3H-indazolyl, an optionally substitutedindolizinyl, an optionally substituted quinolinyl, an optionallysubstituted isoquinolinyl, an optionally substituted benzoxazolyl, anoptionally substituted benzo[1,3]dioxolyl, an optionally substitutedbenzofuryl, an optionally substituted benzothiazolyl, an optionallysubstituted benzo[d]isoxazolyl, an optionally substitutedbenzo[d]isothiazolyl, an optionally substitutedthiazolo[4,5-c]pyridinyl, an optionally substitutedthiazolo[5,4-c]pyridinyl, an optionally substitutedthiazolo[4,5-b]pyridinyl, an optionally substitutedthiazolo[5,4-b]pyridinyl, an optionally substitutedoxazolo[4,5-c]pyridinyl, an optionally substitutedoxazolo[5,4-c]pyridinyl, an optionally substitutedoxazolo[4,5-b]pyridinyl, an optionally substitutedoxazolo[5,4-b]pyridinyl, an optionally substituted imidazopyridinyl, anoptionally substituted benzothiadiazolyl, benzoxadiazolyl, an optionallysubstituted benzotriazolyl, an optionally substituted tetrahydroindolyl,an optionally substituted azaindolyl, an optionally substitutedquinazolinyl, an optionally substituted purinyl, an optionallysubstituted imidazo[4,5-a]pyridinyl, an optionally substitutedimidazo[1,2-a]pyridinyl, an optionally substituted3H-imidazo[4,5-b]pyridinyl, an optionally substituted1H-imidazo[4,5-b]pyridinyl, an optionally substituted1H-imidazo[4,5-c]pyridinyl, an optionally substituted3H-imidazo[4,5-c]pyridinyl, an optionally substituted pyridopyrdazinyl,and optionally substituted pyridopyrimidinyl, an optionally substitutedpyrrolo[2,3]pyrimidyl, an optionally substituted pyrazolo[3,4]pyrimidylan optionally substituted cyclopentaimidazolyl, an optionallysubstituted cyclopentatriazolyl, an optionally substitutedpyrrolopyrazolyl, an optionally substituted pyrroloimidazolyl, anoptionally substituted pyrrolotriazolyl, or an optionally substitutedbenzo(b)thienyl.

In one embodiment, of the compounds represented by formula (I), (II),(III), or(IV), R₅ is selected from the group consisting of:

-   -   wherein:    -   X₁₁, for each occurrence, is independently CH, CR₉, N,N(O), or        N⁺(R₁₇);    -   X₁₂, for each occurrence, is independently CH, CR₉, N,N(O),        N⁺(R₁₇), provided that at least one X₁₂ group is independently        selected from CH and CR₉;    -   X₁₃, for each occurrence, is independently O, S, S(O)_(p), NR₇,        or NR₁₇;    -   R₉, for each occurrence, is independently a substituent selected        from the group consisting of an optionally substituted alkyl, an        optionally substituted alkenyl, an optionally substituted        alkynyl, an optionally substituted cycloalkyl, an optionally        substituted cycloalkenyl, an optionally substituted        heterocyclyl, an optionally substituted aryl, an optionally        substituted heteroaryl, an optionally substituted aralkyl, an        optionally substituted heteraralkyl, halo, cyano, nitro,        guanadino, a hydroxyalkyl, alkoxyalkyl, haloalkyl, a        heteroalkyl, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇,        —C(O)NR₁₀R₁₁, —NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇,        —S(O)_(p)OR₇, —NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇,        —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or        —SP(O)(OR₇)₂;        or two R₉ groups taken together with the carbon atoms to which        they are attached form a fused ring; and    -   R₁₇, for each occurrence, is independently an alkyl or an        aralkyl.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is

In one aspect, R₉ is an optionally substituted heterocyclyl.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is

-   -   wherein R₂₇, for each occurrence, is independently a substituent        selected from the group consisting of —H, an optionally        substituted alkyl, an optionally substituted alkenyl, an        optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, an optionally substituted heteraralkyl,        halo, cyano, nitro, guanadino, a hydroxyalkyl, alkoxyalkyl,        haloalkyl, a heteroalkyl, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇,        —OC(O)R₇, —C(O)NR₁₀R₁₁, —NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇,        —OS(O)_(p)R₇, —S(O)_(p)OR₇, —NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁,        —S(O)_(p)OR₇, —OP(O)(OR₇)₂, —SP(O)(OR₇)₂, —S(O)_(p)OR₇,        —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂;        or two R₂₇ groups taken together with the carbon atom to which        they are attached form an optionally substituted cycloalkyl or        optionally substituted heterocyclyl ring.

In one embodiment, of the compounds represented by formula (I), (II),(III), or (IV), R₅ is X₂₀R₅₀. In one aspect, X₂₀ is a C1-C4 alkyl andR₅₀ is an optionally substituted phenyl.

In one embodiment, of the compounds represented by formula (I), (II), or(III), R₁ and R₃ are each independently —OH, —SH, or —NHR₇. In oneaspect, R₁ and R₃ are both —OH. In one aspect, R₁ is —OH. In one aspect,R₁ is —SH.

In one embodiment, of the compounds represented by formula (I) or (II),one of R₁, R₂, or R₃ is —OP(O)(OH)₂.

In one embodiment, of the compounds represented by formula (I) or (II),two of R₁, R₂, or R₃ are —OP(O)(OH)₂.

In one embodiment, of the compounds represented by formula (I) or (II),all of R₁; R₂, and R₃ are —OP(O)(OH)₂.

In one embodiment, of the compounds represented by formula (I), Z, foreach occurrence, is independently an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl; an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, an optionally substituted heteroaralkyl, halo,cyano, nitro, guanadino, a haloalkyl, a heteroalkyl, alkoxy, haloalkoxy,—NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —C(S)R₇, —C(O)SR₇, —C(S)SR₇,—C(S)OR₇, —C(S)NR₁₀OR₁₁, —C(NR₈)OR₇, —C(NR₈)R₇, —C(NR₈)NR₁₀R₁₁,—C(NR₈)SR₇, —OC(O)R₇, —OC(O)OR₇, —OC(S)OR₇, —OC(NR₈)OR₇, —SC(O)R₇,—SC(O)OR₇, —SC(NR₈)OR₇, —OC(S)R₇, —SC(S)R₇, —SC(S)OR₇, —OC(O)NR₁₀R₁₁,—OC(S)NR₁₀R₁₁, —OC(NR₈)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁, —SC(NR₈)NR₁₀R₁₁,—SC(S)NR₁₀OR₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇, —C(O)NR₁₀R₁₁, —NR₈C(O)R₇,—NR₇C(S)R₇, —NR₇C(S)OR₇, —NR₇C(NR₈)R₇, —NR₇C(O)OR₇, —NR₇C(NR₈)OR₇,—NR₇C(O)NR₁₀R₁₁, —NR₇C(S)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —SR₇, —S(O)_(p)R₇,—OS(O)_(p)R₇, —OS(O)_(p)OR₇, —OS(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇,—NR₈S(O)_(p)R₇, —NR₇S(O)_(p)NR₁₀OR₁₁, —NR₇S(O)_(p)OR₇, —S(O)_(p)NR₁₀R₁₁,—SS(O)_(p)R₇, —SS(O)_(p)OR₇, —SS(O)_(p)NR₁₀R₁₁, —OP(O)(OR₇)₂, or—SP(O)(OR₇)₂. In one aspect, Z is a C1-C6 alkyl, a C1-C6 haloalkyl, aC1-C6 alkoxy, a C1-C6 haloalkoxy, a C₁-C₆ alkyl sulfanyl or a C3-C6cycloalkyl. In one aspect, Z is selected from the group consisting ofmethyl, ethyl, isopropyl, and cyclopropyl.

In one embodiment, of the compounds represented by formula (I), n is 1.

In one embodiment, of the compounds represented by formula (I), n is 0.

In one embodiment, of the compounds represented by formula (I), n is 2.

In one embodiment, of the compounds represented by formula (II), (III),or (IV), R₆ is a C1-C6 alkyl, a C1-C6 haloalkyl, a C1-C6 alkoxy, a C1-C6haloalkoxy, a C1-C6 alkyl sulfanyl or a C3-C6 cycloalkyl. In one aspect,R₆ is selected from the group consisting of methyl, ethyl, isopropyl,and cyclopropyl.

In one embodiment, of the compounds represented by formula (V), (VI), or(VII), R₄₁ is selected from the group consisting of lower alkyl, loweralkoxy, lower cycloalkyl, and lower cycloalkoxy. In one aspect, R₄₁ isselected from the group consisting of methyl, ethyl, propyl, isopropyl,cyclopropyl, methoxy, ethoxy, propoxy, and cyclopropoxy.

In one embodiment, of the compounds represented by formula (V), X₄₁ isNR₄₂ and X₄₂ is CR₄₄.

In one embodiment, of the compounds represented by formula (V), X₄₁ isNR₄₂ and X₄₂ is N.

In one embodiment, of the compounds represented by formula (V), X₄₁ isNR₄₂, and R₄₂ is selected from the group consisting of —H, a loweralkyl, a lower cycloalkyl, —C(O)N(R₂₇)₂, and —C(O)OH, wherein each R₂₇is independently —H or a lower alkyl.

In one embodiment, of the compounds represented by formula (V), X₄₁ isNR₄₂, and R₄₂ is selected from the group consisting of —H, methyl,ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, sec-butyl, tert-butyl,n-pentyl, n-hexyl, —C(O)OH, —(CH₂)_(m)C(O)OH, —CH₂OCH₃, —CH₂CH₂OCH₃, and—C(O)N(CH₃)₂.

In one embodiment, of the compounds represented by formula (V), R₄₃ andR₄₄ are, independently, selected from the group consisting of —H,methyl, ethyl, propyl, isopropyl, cyclopropyl, methoxy, ethoxy, propoxy,and cyclopropoxy.

In one embodiment, of the compounds represented by formula (V), X₄₂ isCR₄₄; Y₄₀ is CR₄₃; and R₄₃ and R₄₄ together with the carbon atoms towhich they are attached form a cycloalkenyl, an aryl, heterocyclyl, orheteroaryl ring.

In one embodiment, of the compounds represented by formula (V), R₄₃ andR₄₄ together with the carbon atoms to which they are attached form aC₅-C₈ cycloalkenyl or a C₅-C₈ aryl.

In one embodiment, of the compounds represented by formula (V), R₄₅ orCR₄₅ is selected from the group consisting of —H, —OH, —SH, —NH₂, alower alkoxy, a lower alkyl amino, and a lower dialkyl amino.

In one embodiment, of the compounds represented by formula (V), R₄₅ isselected from the group consisting of —H, —OH, methoxy and ethoxy.

In one embodiment, of the compounds represented by formula (V), X₄₁ isO.

In one embodiment, of the compounds represented by formula (VI), X₄₂ isCR₄₄, and R₄₃ and R₄₄ are, independently, selected from the groupconsisting of —H, methyl, ethyl, propyl, isopropyl, cyclopropyl,methoxy, ethoxy, propoxy, and cyclopropoxy.

In one embodiment, of the compounds represented by formula (VI), X₄₂ isCR₄₄, and R₄₃ and R₄₄, taken together with the carbon atoms to whichthey are attached, form a cycloalkenyl, aryl, heterocyclyl, orheteroaryl ring.

In one embodiment, of the compounds represented by formula (VI), R₄₃ andR₄₄, taken together with the carbon atoms to which they are attached,form a C₅-C₈ cycloalkenyl or a C₅-C₈ aryl.

In one embodiment, of the compounds represented by formula (VI), X₄₂ isCR₄₄.

In one embodiment, of the compounds represented by formula (VI), X₄₂ isN.

In one embodiment, of the compounds represented by formula (VIII), R₄₂is —H or an optionally substituted lower alkyl.

In another embodiment, the compound is selected from the groupconsisting of

-   4-(4-(2,3-dihydro-1H-inden-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyl    dihydrogen phosphate;-   5-hydroxy-4-(5-hydroxy-4-(6-morpholinopyridin-3-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyl    dihydrogen phosphate;-   5-hydroxy-4-(5-hydroxy-4-(1-methyl-1H-indol-5-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyl    dihydrogen phosphate;-   sodium    5-hydroxy-4-(5-hydroxy-4-(1-methyl-1H-indol-5-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyl    phosphate;-   2-(4-(2,3-dihydro-1H-inden-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-4-isopropylphenyl    dihydrogen phosphate;-   4-(2,3-dihydro-1H-inden-5-yl)-5-(2,4-dihydroxy-5-isopropylphenyl)-4H-1,2,4-triazol-3-yldihydrogen    phosphate;-   4-(4-(1′,3′-dihydrospiro[[1,3]dioxolane-2,2′-indene]-5′-yl)-5-mercapto-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyl    dihydrogen phosphate;-   2-(3,4-dimethoxyphenethyl)-5-hydroxy-4-(5-hydroxy-4-(1-methyl-1H-indol-5-yl)-4H-1,2,4-triazol-3-yl)phenyl    dihydrogen phosphate;-   4-(4-(2,3-dihydro-1H-inden-5-yl)-5-(phenylamino)-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyl    dihydrogen phosphate;-   5-hydroxy-2-isopropyl-4-(5-mercapto-4-(4-methoxybenzyl)-4H-1,2,4-triazol-3-yl)phenyl    dihydrogen phosphate;-   5-hydroxy-4-(5-hydroxy-4-(4-methoxybenzyl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyl    dihydrogen phosphate;-   4-(4-((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)methyl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyl    dihydrogen phosphate;-   4-(4-(4-bromo-2-methylphenyl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-3-hydroxyphenyl    dihydrogen phosphate; or-   4-(4-(1,3-dimethyl-1H-indol-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-2-ethyl-5-hydroxyphenyl    dihydrogen phosphate;    -   or a tautomer, pharmaceutically acceptable salt, solvate,        clathrate, or a prodrug thereof.

In one embodiment, the compounds of the invention do not include thecompounds disclosed in U.S. patent application Ser. No. 11/282,119,filed Nov. 17, 2005.

Exemplary compounds of the invention are depicted in Table 1 below,including tautomers, pharmaceutically acceptable salts, solvates,clathrates, hydrates, polymorphs or prodrugs thereof.

NO. Structure Tautomeric structure Name 1

4-(4-(2,3- dihydro-1H- inden-5-yl)-5- hydroxy-4H- 1,2,4-triazol-3-yl)-5-hydroxy-2- isopropylphenyl dihydrogen phosphate 2

5-hydroxy-4-(5- hydroxy-4-(6- morpho- linopyridin- 3-yl)-4H-1,2,4-triazol-3-yl)-2- isopropylphenyl dihydrogen phosphate 3

5-hydroxy-4-(5- hydroxy-4-(1- methyl-1H-indol- 5-yl)-4H-1,2,4-triazol-3-yl)-2- isopropylphenyl dihydrogen phosphate 4

sodium 5- hydroxy-4-(5- hydroxy-4-(1- methyl-1H-indol- 5-yl)-4H-1,2,4-triazol-3-yl)-2- isopropylphenyl phosphate 5

2-(4-(2,3- dihydro-1H- inden-5-yl)-5- hydroxy-4H- 1,2,4-triazol-3-yl)-5-hydroxy-4- isopropylphenyl dihydrogen phosphate 6

4-(2,3-dihydro- 1H-inden-5-yl)- 5-(2,4- dihydroxy-5- isopropylphenyl)-4H-1,2,4-triazol- 3-yl dihydrogen phosphate 7

4-(4-(1′,3′- dihydrospiro[[1,3] dioxolane-2,2′- indene]-5′-yl)-5-mercapto-4H- 1,2,4-triazol-3-yl)- 5-hydroxy-2- isopropylphenyldihydrogen phosphate 8

2-(3,4- dimethoxyphen- ethyl)-5-hydroxy- 4-(5-hydroxy-4- (1-methyl-1H-indol-5-yl)-4H- 1,2,4-triazol-3-yl) phenyl dihydrogen phosphate 9

4-(4-(2,3- dihydro-1H- inden-5-yl)-5- (phenylamino)- 4H-1,2,4-triazol-3-yl)-5-hydroxy- 2-isopropylphenyl dihydrogen phosphate 10

5-hydroxy-2- isopropyl-4-(5- mercapto-4-(4- methoxybenzyl)-4H-1,2,4-triazol- 3-yl)phenyl dihydrogen phosphate 11

5-hydroxy-4-(5- hydroxy-4-(4- methoxybenzyl)- 4H-1,2,4-triazol- 3-yl)-2-isopropylphenyl dihydrogen phosphate 12

4-(4-((2,3- dihydrobenzo[b] [1,4]dioxin-6-yl) methyl)-5- hydroxy-4H-1,2,4-triazol-3- yl)-5-hydroxy-2- isopropylphenyl dihydrogen phosphate13

4-(4-(4-bromo-2- methylphenyl)-5- hydroxy-4H- 1,2,4-triazol-3-yl)-3-hydroxyphenyl dihydrogen phosphate 14

4-(4-(1,3- dimethyl-1H- indol-5-yl)-5- hydroxy-4H- 1,2,4-triazol-3-yl)-2-ethyl-5- hydroxyphenyl dihydrogen phosphate

In certain instances tautomeric forms of the disclosed compound exist,such as the tautomeric structures shown below:

X₁₅═O,S or NR₇

It is to be understood that when a compound is represented by astructural formula herein, all other tautomeric forms which may existfor the compound are encompassed the structural formula. Compoundsrepresented by formulas disclosed herein that can form analogoustautomeric structures to the one shown above are also preferred.

Without wishing to be bound by any theory, it is believed that thecompounds of the invention preferentially bind to Hsp90 in thetautomeric form shown above, and thereby inhibit the activity of Hsp90.

C. Methods for Making Compounds of the Invention

Compounds of the invention can be obtained via standard, well-knownsynthetic methodology, see e.g., March, J. Advanced Organic Chemistry;Reactions Mechanisms, and Structure, 4th ed., 1992. In particular,compounds of the invention can be obtained by heating a benzoic acid (1)with an aminophenyl (2) to give a phenyl benzamide (3) which can then bereacted with hydrazine to give a triazole (4) (see Scheme I below).Starting materials useful for preparing compounds of the invention andintermediates therefore, are commercially available or can be preparedfrom commercially available materials using known synthetic methods andreagents.

Additional methods of preparing the compounds of the invention can befound in U.S. patent application Ser. No. 11/807,333, U.S. patentapplication Ser. No. 11/807,331, U.S. patent application Ser. No.11/807,327, and U.S. patent application Ser. No. 11/807,201, the entireteachings of each of these applications are incorporated herein byreference. Reactive functional groups can be protected during one ormore reaction step, then deprotected to restore the originalfunctionality. Examples of suitable protecting groups for hydroxylgroups include benzyl, methoxymethyl, allyl, trimethylsilyl,tert-butyldimethylsilyl, acetate, and the like. Examples of suitableamine protecting groups include benzyloxycarbonyl, tert-butoxycarbonyl,tert-butyl, benzyl and fluorenylmethyloxy-carbonyl (Fmoc). Examples ofsuitable thiol protecting groups include benzyl, tert-butyl, acetyl,methoxymethyl and the like. Other suitable protecting groups are wellknown to those of ordinary skill in the art and include those found inT. W. Greene, Protecting Groups in Organic Synthesis, John Wiley & Sons,Inc. 1981.

In addition, compounds of the invention can also be prepared as shownbelow in the Schemes and Examples below.

In one embodiment, the compounds can be prepared as shown in Scheme II.A dihydroxy benzoic acid methyl ester is reacted with benzyl chloride,to produce a Bis-benzyloxy benzoic acid methyl ester (1). TheBis-benzyloxy benzoic acid methyl ester can then be heated with LiOH togive a Bis-benzyloxy benzoic acid (2). The Bis-benzyloxy benzoic acid(2) is then reacted with an aminophenyl to produce a phenyl-benzamide(3). The phenyl-benzamide (3) is then reacted with hydrazine to give atriazol (4). The hydroxy groups can then be unprotected in the presenceof palladium on charcoal to give the final product.

In another embodiment the compounds can be prepared as shown in SchemeIII. A nitroaniline (1) can be reacted with propionyl chloride to yieldnitro-phenyl-propionamide (2). NaH can then be added to a solution of(2) in anhydrous THF followed by iodomethane to give pure productnitro-phenyl-N-methyl-propionamide (3).

The nitro-phenyl-N-methyl-propionamide (3) and borane-methyl sulfidecomplex are heated to give the nitro-phenyl-methyl-propyl-amine (4). Asolution of (4) in MeOH/EtOAc containing Pd—C can be subjected tohydrogenation to give the N—methyl-N-propyl-benzene-1,3-diamine (5).

To a stirred solution of (5) in CH₂Cl₂ can be added1,1′-thiocarbonyldiimidazole to give the(5-Isothiocyanato-2-methoxy-phenyl)-methyl-propyl-amine (6).

The isothiocyanate (6) can be reacted with the hydrazide (7) to give theintermediate (8). A solution of NaOH in water can be added to theintermediate (8), which can then be flushed with nitrogen and heated.The reaction mixture can then be cooled and acidified. The mixture canthen be filtered and purified to give4-isopropyl-6-{5-mercapto-4-[4-methoxy-3-(methyl-propyl-amino)-phenyl]-4H[1,2,4.]triazol-3-yl}-benzene-1,3-diol.

In another embodiment the compounds can be prepared as shown in SchemeIV. A bromo-nitrobenzene (1) can be reacted with N¹, N², N²trimethylethane-1,2-diamine to give N¹-(nitrophenyl)-N¹, N²,N²-trimethylethane-1,2-diamine (2). A solution of (2) in can besubjected to hydrogenation, passed through a short pad of celite, washedwith MeOH and evaporated under reduced pressure. Thiocarbodiimidazolecan then be added to (2) to give the N¹-(isothiocyanato-phenyl)-)-N¹,N², N²-trimethylethane-1,2-diamine (3).

The isothiocyanate (3) can then be reacted with a benzoic acid hydrazideto give the final product4-(4-(3-((2-(dimethylamino)ethyl)(methyl)amino)-4-methoxyphenyl)-5-mercapto-4H-1,2,4-triazol-3-yl)-6-isopropylbenzene-1,3-diol(4).

D. Uses of Compounds of the Invention

The present invention is directed to therapies which involveadministering one of more compounds of the invention, and compositionscomprising said compounds to a subject, preferably a human subject, toinhibit the activity of Hsp90 or to prevent, treat, manage, orameliorate a proliferative disorder, such as cancer, or one or moresymptoms thereof.

In one embodiment, the present invention is directed to treating cancersin which aberrant expression and/or activation of c-kit has beenimplicated as a contributing factor. The method comprises administeringto a patient an effective amount of a compound represented by formula(I)-(VIII), or any embodiment thereof, or a compound shown in Table 1.

In one embodiment, the present invention is directed to treating cancersin which expression of Bcr-Abl has been implicated as a contributingfactor. The method comprises administering to a patient an effectiveamount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1.

In one embodiment, the present invention is directed to treating cancersin which aberrant expression and/or activation of flt-3 has beenimplicated as a contributing factor. The method comprises administeringto a patient an effective amount of a compound represented by formula(I)-(VIII), or any embodiment thereof, or a compound shown in Table 1.

In one embodiment, the present invention is directed to treating cancersin which aberrant expression and/or activation of EGFR has beenimplicated as a contributing factor. The method comprises administeringto a patient an effective amount of a compound represented by formula(I)-(VIII), or any embodiment thereof, or a compound shown in Table 1.

In one embodiment, the present invention is directed to treating cancersin which Hsp90 is over expressed compared with normal cells. The methodcomprises administering to a patient an effective amount of a compoundrepresented by formula (I)-(VIII), or any embodiment thereof, or acompound shown in Table 1. Examples of cancers in which Hsp90 is overexpressed include difuse large B-cell lymphomas (DLBCL).

In one aspect, the invention provides a method of inhibiting theactivity of Hsp90 in a cell, comprising administering to the cell aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the compound is administered to a cell in a subject, preferably amammal, and more preferably a human.

In another aspect, the invention provides a method of treating orpreventing a proliferation disorder in a mammal, comprisingadministering to the mammal an effective amount of a compoundrepresented by formula (I)-(VIII), or any embodiment thereof, or acompound shown in Table 1. In one embodiment, the compound isadministered to a human to treat or prevent a proliferative disorder. Inanother embodiment, the proliferation disorder is cancer. In anotherembodiment, the compound is administered with one or more additionaltherapeutic agents. In a preferred embodiment, the additionaltherapeutic agent is an anticancer agent.

In another aspect, the invention provides a method for treating cancerin a mammal, comprising administering to the mammal an effective amountof a compound represented by formula (I)-(VIII), or any embodimentthereof, or a compound shown in Table 1. In one embodiment, the compoundis administered to a human to treat or prevent cancer. In anotherembodiment, the compound is administered with one or more additionaltherapeutic agents. In a preferred embodiment, the one or moreadditional therapeutic agents are anticancer agents.

In another aspect, the invention provides a method for treating a c-kitassociated cancer in a mammal, comprising administering to the mammal aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the compound is administered to a human to treat or prevent the c-kitassociated cancer. In another embodiment, the compound is administeredwith one or more additional therapeutic agents. In a preferredembodiment, the one or more additional therapeutic agents are anticanceragents.

In another aspect, the invention provides a method for treating aBcr-Abl associated cancer in a mammal, comprising administering to themammal an effective amount of a compound represented by formula(I)-(VIII), or any embodiment thereof, or a compound shown in Table 1.In one embodiment, the compound is administered to a human to treat orprevent the Bcr-Abl associated cancer. In another embodiment, thecompound is administered with one or more additional therapeutic agents.In a preferred embodiment, the one or more additional therapeutic agentsare anticancer agents.

In another aspect, the invention provides a method for treating a flt3associated cancer in a mammal, comprising administering to the mammal aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the compound is administered to a human to treat or prevent the flt3associated cancer. In another embodiment, the compound is administeredwith one or more additional therapeutic agents. In a preferredembodiment, the one or more additional therapeutic agents are anticanceragents.

In another aspect, the invention provides a method for treating an EGFRassociated cancer in a mammal, comprising administering to the mammal aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the compound is administered to a human to treat or prevent the EGFRassociated cancer. In another embodiment, the compound is administeredwith one or more additional therapeutic agents. In a preferredembodiment, the one or more additional therapeutic agents are anticanceragents.

In another aspect, the invention provides a method for treating a cancerin a mammal which is characterized by the upregulation of Hsp90 comparedto normal cells of the same type, comprising administering to the mammalan effective amount of a compound represented by formula (I)-(VIII), orany embodiment thereof, or a compound shown in Table 1. In oneembodiment, the compound is administered to a human to treat or preventthe cancer associated with the upregulation of Hsp90. In anotherembodiment, the cancer associated with the upregulation of Hsp90 isDLBCL. In another embodiment, the compound is administered with one ormore additional therapeutic agents. In a preferred embodiment, the oneor more additional therapeutic agents are anticancer agents.

In another aspect, the invention provides a method for treating orinhibiting angiogenesis in a subject in need thereof, comprisingadministering to the subject an effective amount of a compoundrepresented by formula (I)-(VIII), or any embodiment thereof, or acompound shown in Table 1.

In another aspect, the invention provides a method of blocking,occluding, or otherwise disrupting blood flow in neovasculature,comprising contacting the neovasculature with an effective amount of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1. In one aspect, the neovasculature is ina subject and blood flow in the neovasculature is blocked, occluded, orotherwise disrupted in the subject by administering to the subject aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one aspect, thesubject is human.

The present invention provides a method for preventing, treating,managing, or ameliorating an infection in a subject in need thereof,comprising administering an effective amount of a compound representedby formula (I)-(VIII), or any embodiment thereof, or a compound shown inTable 1.

In one aspect, the invention is directed to a method of treating orpreventing a fungal infection.

In one aspect, the invention is directed to a method of treating orpreventing a yeast infection.

In one aspect, the invention is directed to a method of treating orpreventing a yeast infection caused by a Candida yeast.

In another embodiment the invention is directed to a method of treatingor preventing fungal drug resistance. In one aspect, the fungal drugresistance is associated with an azole drug. In another aspect, thefungal drug resistance is associated with a non-azole fungal drug. Inone aspect, the non-azole drug is an echinocandin. In one aspect, theazole fungal drug is ketoconazole, miconazole, fluconazole,itraconazole, posaconazole, ravuconazole, voriconazole, clotrimazole,econazole, oxiconazole, sulconazole, terconazole, butoconazole,isavuconazole, or tioconazole. In one aspect, the azole fugnal drug isfluconazole.

In one aspect, the invention is directed to a method of treating orpreventing a bacterial infection.

In one aspect, the invention is directed to a method of treating orpreventing a bacterial infection caused by a Gram Positive Bacteria.

In one aspect, the invention is directed to a method of treating orpreventing a bacterial infection caused by a Gram Negative Bacteria.

In one aspect, the invention is directed to a method of treating orpreventing a viral infection.

In one aspect, the invention is directed to a method of treating orpreventing a viral infection caused by an influenza virus, a herpesvirus, a hepatitis virus, or an HIV virus.

In one aspect, the invention is directed to a method of treating orpreventing a viral infection caused by influenza A virus, herpes simplexvirus type 1, hepatitis C virus, hepatitis B virus, HIV-1 virus, orEpstein-Barr Virus.

In one aspect, the invention is directed to a method of treating orpreventing a parasitic infection.

In one aspect, the invention is directed to a method of treating orpreventing a protozoal infection.

In one aspect, the invention is directed to a method of treating orpreventing an infection caused by plasmodium falciparum or trypsanosomacruzi.

In one aspect, the invention is directed to a method of treating orpreventing an infection caused by a leishmania protozoa.

In one aspect, the invention is directed to a method of treating orpreventing an amoebic infection.

In one aspect, the invention is directed to a method of treating orpreventing a helminth infection.

In one aspect, the invention is directed to a method of treating orpreventing an infection caused by schistostoma mansoni.

In one aspect, compounds of the invention are administered incombination with one or more additional anti-infective therapeuticagents.

The present invention provides a method for inhibiting topoisomerase II,comprising administering an effective amount of a compound representedby formula (I)-(VIII), or any embodiment thereof, or a compound shown inTable 1.

In another embodiment, topoisomerase II is associated with a disease andadministering the compound will treat or prevent the disease.

In one aspect, the disease is a proliferative disease.

In another aspect, the proliferative disease is cancer.

In one aspect, the disease is an infection.

The present invention provides a method of treating an inflammatorydisorder in a subject in need thereof, comprising administering aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the inflammatory disorder is selected from the group consisting oftransplant rejection, skin graft rejection, arthritis, rheumatoidarthritis, osteoarthritis and bone diseases associated with increasedbone resorption; inflammatory bowel disease, ileitis, ulcerativecolitis, Barrett's syndrome, Crohn's disease; asthma, adult respiratorydistress syndrome, chronic obstructive airway disease; cornealdystrophy, trachoma, onchocerciasis, uveitis, sympathetic ophthalmitis,endophthalmitis; gingivitis, periodontitis; tuberculosis; leprosy;uremic complications, glomerulonephritis, nephrosis; sclerodermatitis,psoriasis, eczema; chronic demyelinating diseases of the nervous system,multiple sclerosis, AIDS-related neurodegeneration, Alzheimer's disease,infectious meningitis, encephalomyelitis, Parkinson's disease,Huntington's disease, amyotrophic lateral sclerosis viral or autoimmuneencephalitis; autoimmune disorders, immune-complex vasculitis, systemiclupus and erythematodes; systemic lupus erythematosus (SLE);cardiomyopathy, ischemic heart disease hypercholesterolemia,atherosclerosis, preeclampsia; chronic liver failure, brain and spinalcord trauma.

The present invention provides a method of treating an immune disorderin a subject in need thereof, comprising administering an effectiveamount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the immune disorder is selected from the group consisting of multiplesclerosis, myasthenia gravis, Guillain-Barré, autoimmune uveitis,autoimmune hemolytic anemia, pernicious anemia, autoimmunethrombocytopenia, temporal arteritis, anti-phospholipid syndrome,vasculitides such as Wegener's granulomatosis, Behcet's disease,psoriasis, dermatitis herpetiformis, pemphigus vulgaris, vitiligo,Crohn's disease, ulcerative colitis, primary biliary cirrhosis,autoimmune hepatitis, Type 1 or immune-mediated diabetes mellitus,Grave's disease. Hashimoto's thyroiditis, autoimmune oophoritis andorchitis, autoimmune disorder of the adrenal gland, rheumatoidarthritis, systemic lupus erythematosus, scleroderma, polymyositis,dermatomyositis, ankylosing spondylitis, and Sjogren's syndrome.

The present invention provides a method of suppressing an immuneresponse in a subject in need thereof, comprising administering aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one embodiment,the subject in need of immunosuppression is a subject that has receivedan organ or tissue transplant, such as a skin graft, heart, kidney,lung, liver, pancreas, cornea, bowel, stomach, and the like. In anotherembodiment, the subject in need of immunosuppression is a subject thathas received stem cell transplantation. The transplant may be asyngeneic transplant (i.e., from a donor that has the same genetic makeup), an allographic transplant (i.e., from a donor of the same species)or a xenographic transplant (i.e., from a donor that is a differentspecies). The present invention provides a method of inhibiting theproduction of inflammatory cytokines, such as G-CSF, GM-CSF, IL-12,IL-1β, IL-23, IL-6, IL-8, and TNF-α, in a subject in need of suchtreatment. The method comprises administering to the subject aneffective amount of a compound represented by formula (I)-(VIII), or anyembodiment thereof, or a compound shown in Table 1. In one aspect, thedosage solution is administered intravenously.

The present invention provides a dosing solution containing 1 mg/mL of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1 in 15 mM phosphate buffer with a pH inthe range of 5 to 7. In one aspect, the dosing solution has anosmolarity of about 353 mOsm/kg to 390 mOsm/kg. In one aspect, thedosing solution has an osmolarity of about 353 mOsm/kg. In one aspect,the dosing solution has a pH of about 7. In one aspect, the dosagesolution is administered intravenously.

The present invention provides a dosing solution containing 6 mg/mL of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1 in 15 mM phosphate buffer with a pH inthe range of 5 to 7. In one aspect, the dosing solution has anosmolarity of about 353 mOsm/kg to 390 mOsm/kg. In one aspect, thedosing solution has an osmolarity of about 390 mOsm/kg. In one aspect,the dosing solution has a pH of about 5. In one aspect, the dosagesolution is administered intravenously.

The present invention provides a dosing solution containing 3 mg/mL of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1 in Plasmalyte 148 Injection USP with a pHin the range of 5 to 7. In one aspect, the pH is adjusted with NaOH.

The present invention provides a method for the administration of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1 to a mammal comprising employing a dosingsolution containing 1 mg/mL of the compound in 15 mM phosphate bufferwith a pH in the range of 5 to 7. In one aspect, the dosing solution hasan osmolarity of about 353 mOsm/kg to 390 mOsm/kg. In one aspect, thedosing solution has an osmolarity of about 353 mOsm/kg. In one aspect,the dosing solution has a pH of about 7.

The present invention provides a method for the administration of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1 to a mammal comprising employing a dosingsolution containing 6 mg/mL of the compound in 15 mM phosphate bufferwith a pH in the range of 5 to 7. In one aspect, the dosing solution hasan osmolarity of about 353 mOsm/kg to 390 mOsm/kg. In one aspect, thedosing solution has an osmolarity of about 390 mOsm/kg. In one aspect,the dosing solution has a pH of about 5.

The present invention provides a method for the administration of acompound represented by formula (I)-(VIII), or any embodiment thereof,or a compound shown in Table 1 to a mammal comprising employing a dosingsolution containing 3 mg/mL of the compound in Plasmalyte 148 InjectionUSP with a pH in the range of 5 to 7. In one aspect, the pH is adjustedwith NaOH.

1. c-Kit Associated Cancers

SCF binding to the c-kit protects hematopoietic stem and progenitorcells from apoptosis (Lee, et al., 1997, J. Immunol., 159:3211-3219),thereby contributing to colony formation and hematopoiesis. Expressionof c-kit is frequently observed in acute myelocytic leukemia (AML) andsometimes observed in acute lymphocytic leukemia (ALL) (for reviews, seeSperling, et al., 1997, Haemat., 82:617-621; Escribano, et al., 1998,Leuk. Lymph., 30:459-466). Although c-kit is expressed in the majorityof AML cells, its expression does not appear to be prognostic of diseaseprogression (Sperling, et al, 1997, Haemat. 82:617-621). However, SCFprotected AML cells from apoptosis induced by chemotherapeutic agents(Hassan, et al., 1996, Acta. Hem., 95:257-262). Therefore, degradationof c-kit caused by the inhibition of Hsp90 by the compounds of theinvention will enhance the efficacy of these agents and may induceapoptosis of AML cells.

The clonal growth of cells from patients with myelodysplastic syndrome(Sawada, et al., 1996, Blood, 88:319-327) or chronic myelogenousleukemia (CML) (Sawai, et al., 1996, Exp. Hem., 2:116-122) was found tobe significantly enhanced by SCF in combination with other cytokines.CML is characterized by expansion of Philadelphia chromosome positivecells of the marrow (Verfaillie, et al., 1998, Leuk., 12:136-138), whichappears to primarily result from inhibition of apoptotic death (Jones,1997, Curr. Opin. One., 9:3-7). The product of the Philadelphiachromosome, p210.sup.BCR-ABL, has been reported to mediate inhibition ofapoptosis (Bedi, et al., 1995, Blood, 86:1148-1158). Sincep210.sup.BCR-ABL and the c-kit RTK both inhibit apoptosis andp62.sup.dok has been suggested as a substrate (Carpino, et al., 1997,Cell, 88:197-204), it is possible that clonal expansion mediated bythese kinases occurs through a common signaling pathway. However, c-kithas also been reported to interact directly with p210.sup.BCR-ABL(Hallek, et al., 1996, Brit. J Haem., 94:5-16), which suggests thatc-kit may have a more causative role in CML pathology. Therefore,degradation of c-kit caused by the inhibition of Hsp90 by the compoundsof the invention will prove useful in the treatment of CML.

Normal colorectal mucosa does not express c-kit (Bellone, et al., 1997,J. Cell Physiol., 172:1-11). However, c-kit is frequently expressed incolorectal carcinoma (Bellone, et al., 1997, J. Cell Physiol., 172:1-11), and autocrine loops of SCF and c-kit have been observed inseveral colon carcinoma cell lines (Toyota, et al., 1993, Turn. Biol.,14:295-302; Lahm, et al., 1995, Cell Growth & Differ., 6:1111-1118;Bellone, et al., 1997, J. Cell Physiol., 172:1-11). Furthermore,disruption of the autocrine loop by the use of neutralizing antibodies(Lahm, et al., 1995, Cell Growth & Differ., 6:1111-1118) anddownregulation of c-kit and/or SCF significantly inhibits cellproliferation (Lahm, et al., 1995, Cell Growth & Differl., 6:1111-1118;Bellone, et al., 1997, J. Cell Physiol., 172:1-11).

SCF/c-kit autocrine loops have been observed in gastric carcinoma celllines (Turner, et al., 1992, Blood, 80:374-381; Hassan, et al., 1998,Digest. Dis. Science, 43:8-14), and constitutive c-kit activation alsoappears to be important for gastrointestinal stromal tumors (GISTs).GISTs are the most common mesenchymal tumor of the digestive system.More than 90% of GISTs express c-kit, which is consistent with theputative origin of these tumor cells from interstitial cells of Cajal(ICCs) (Hirota, et al., 1998, Science, 279:577-580). The c-kit expressedin GISTs from several different patients was observed to have mutationsin the intracellular juxtamembrane domain leading to constitutiveactivation (Hirota, et al., 1998, Science 279:577-580). Therefore,degradation of c-kit caused by the inhibition of Hsp90 by the compoundsof the invention will be an efficacious means for the treatment of thesecancers.

Male germ cell tumors have been histologically categorized intoseminomas, which retain germ cell characteristics, and nonseminomaswhich can display characteristics of embryonal differentiation. Bothseminomas and nonseminomas are thought to initiate from a preinvasivestage designated carcinoma in situ (CIS) (Murty, et al., 1998, Sem.Oncol., 25:133-144). Both c-kit and SCF have been reported to beessential for normal gonadal development during embryogenesis (Loveland,et al., 1997, J. Endocrinol., 153:337-344). Loss of either the receptoror the ligand resulted in animals devoid of germ cells. In postnataltestes, c-kit has been found to be expressed in Leydig cells andspermatogonia, while SCF was expressed in Sertoli cells (Loveland, etal., 1997, J. Endocrinol., 153:337-344). Testicular tumors develop fromLeydig cells with high frequency in transgenic mice expressing humanpapilloma virus 16 (HPV16) E6 and E7 oncogenes (Kondoh, et al., 1991, J.Virol., 65:3335-3339; Kondoh, et al., 1994, J. Urol., 152:2151-2154).These tumors express both c-kit and SCF, and an autocrine loop maycontribute to the tumorigenesis (Kondoh, et al., 1995, Oncogene,10:341-347) associated with cellular loss of functional p53 and theretinoblastoma gene product by association with E6 and E7 (Dyson, etal., 1989, Science, 243:934-937; Werness, et al., 1990, Science,248:76-79; Scheffner, et al., 1990, Cell, 63:1129-1136). Defectivesignaling mutants of SCF (Kondoh, et al., 1995, Oncogene, 10:341-347) orc-kit (Li, et al., 1996, Canc. Res., 56:4343-4346) inhibited formationof testicular tumors in mice expressing HPV16 E6 and E7. Since c-kitkinase activation is pivotal to tumorigenesis in these animals, thecompounds of the invention which inhibit Hsp90 and thereby cause thedegradation of c-kit will be useful for preventing or treatingtesticular tumors associated with human papilloma virus.

Expression of c-kit on germ cell tumors shows that the receptor isexpressed by the majority of carcinomas in situ and seminomas, but c-kitis expressed in only a minority of nonseminomas (Strohmeyer, et al.,1991, Canc. Res., 51:1811-1816; Rajpert-de Meyts, et al., 1994, Int. J.Androl., 17:85-92; Izquierdo, et al., 1995, J. Pathol., 177:253-258;Strohmeyer, et al., 1995, J. Urol., 153:511-515; Bokenmeyer, et al.,1996, J. Cance. Res., Clin. Oncol., 122:301-306; Sandlow, et al., 1996,J. Androl., 17:403 -408). Therefore, degradation of c-kit caused by theinhibition of Hsp90 by the compounds of the invention will be anefficacious means for the treatment of these cancers.

SCF and c-kit are expressed throughout the central nervous system ofdeveloping rodents, and the pattern of expression suggests a role ingrowth, migration and differentiation of neuroectodermal cells.Expression of SCF and c-kit have also been reported in the adult brain(Hamel, et al., 1997, J. Neuro-Onc., 35:327-333). Expression of c-kithas also been observed in normal human brain tissue (Tada, et al. 1994,J. Neuro., 80:1063-1073). Glioblastoma and astrocytoma, which define themajority of intracranial tumors, arise from neoplastic transformation ofastrocytes (Levin, et al., 1997, Principles & Practice of Oncology,2022-2082). Expression of c-kit has been observed in glioblastoma celllines and tissues (Berdel, et al., 1992, Canc. Res., 52:3498-3502; Tada,et al., 1994, J. Neuro., 80:1063-1073; Stanulla, et al., 1995, Act.Neuropath., 89:158-165).

The association of c-kit with astrocytoma pathology is less clear.Reports of expression of c-kit in normal astrocytes have been made(Natali, et al., 1992, Int. J. Canc., 52:197-201), (Tada, et al. 1994,J. Neuro., 80:1063-1073), while others report it is not expressed(Kristt, et al., 1993, Neuro., 33:106-115). In the former case, highlevels of c-kit expression in high grade tumors were observed (Kristt,et al., 1993, Neuro., 33:106-115), whereas in the latter caseresearchers were unable to detect any expression in astrocytomas. Inaddition, contradictory reports of c-kit and SCF expression inneuroblastomas also exist. One study found that neuroblastoma cell linesoften express SCF, but rarely express c-kit. In primary tumors, c-kitwas detected in about 8% of neuroblastomas, while SCF was found in 18%of tumors (Beck, et al., 1995, Blood, 86:3132-3138). In contrast, otherstudies (Cohen, et al., 1994, Blood, 84:3465-3472) have reported thatall 14 neuroblastoma cell lines examined contained c-kit/SCF autocrineloops, and expression of both the receptor and ligand were observed in45% of tumor samples examined. In two cell lines, anti-c-kit antibodiesinhibited cell proliferation, suggesting that the SCF/c-kit autocrineloop contributed to growth (Cohen, et al., 1994, Blood, 84:3465-3472).Therefore, degradation of c-kit caused by the inhibition of Hsp90 by thecompounds of the invention will be an efficacious means for treatingsome cancers of the central nervous system.

2. Bcr-Abl Associated Cancers

The Philadelphia chromosome which generates the fusion protein Bcr-Ablis associated with the bulk of chronic myelogenous leukemia (CML)patients (more than 95%), 10-25% of acute lymphocytic leukemia (ALL)patients, and about 2-3% of acute myelogenous leukemias (AML). Inaddition, Bcr-Abl is a factor in a variety of other hematologicalmalignancies, including granulocytic hyperplasia resembling CML,myelomonocytic leukemia, lymphomas, and erythroid leukemia (see Lugo, etal., MCB (1989), 9:1263-1270; Daley, et al., Science (1990),247:824-830; and Honda, Blood (1998), 91:2067-2075, the entire teachingsof each of these references are incorporated herein by reference).

A number of different kinds of evidence support the contention thatBcr-Abl oncoproteins, such as p210 and p185 BCR-ABL, are causativefactors in these leukemias (Campbell and Arlinghaus, “Current Status ofBcr Gene Involvement with Human Leukemia”, In: Advances in CancerResearch, Eds. Klein, VandeWoude, Orlando, Fla. Academic Press, Inc.,57:227-256, 1991, the entire teachings of which are incorporated hereinby reference). The malignant activity is due in large part to theBcr-Abl protein's highly activated protein tyrosine kinase activity andits abnormal interaction with protein substrates (Arlinghaus et al., In:UCLA Symposia on Molecular and Cellular Biology New Series, AcuteLymphoblastic Leukemia, Eds. R. P. Gale, D. Hoelzer, New York, N.Y.,Alan R. Liss, Inc., 108:81-90, 1990, the entire teachings of which areincorporated herein by reference). The Bcr-Abl oncoprotein p210 Bcr-Ablis associated with both CML and ALL, whereas the smaller oncoprotein,p185 BCR-ABL, is associated with ALL patients, although some CMLpatients also express p185 (Campbell et al., 1991).

3. FLT3 Associated Cancers

FLT3 associated cancers are cancers in which inappropriate FLT3 activityis detected. FLT3 associated cancers include hematologic malignanciessuch as leukemia and lymphoma. In some embodiments FLT3 associatedcancers include acute myelogenous leukemia (AML), B-precursor cell acutelymphoblastic leukemia, myelodysplastic leukemia, T-cell acutelymphoblastic leukemia, mixed lineage leukemia (MLL), or chronicmyelogenous leukemia (CML).

4. EGFR Associated Cancers

EGFR associated cancers are cancers in which inappropriate EGFR activity(e.g., overexpression of EGFR or mutation of EGFR which causesconstitutive tyrosine kinase activity) has been implicated as acontributing factor. Inappropriate EGFR activity has been associatedwith an adverse prognosis in a number of human cancers, such asneuroblastoma, intestine carcinoma such as rectum carcinoma, coloncarcinoma, familiary adenomatous polyposis carcinoma and hereditarynon-polyposis colorectal cancer, esophageal carcinoma, labial carcinoma,larynx carcinoma, hypopharynx carcinoma, tong carcinoma, salivary glandcarcinoma, gastric carcinoma, adenocarcinoma, medullary thyroideacarcinoma, papillary thyroidea carcinoma, renal carcinoma, kidneyparenchym carcinoma, ovarian carcinoma, cervix carcinoma, uterine corpuscarcinoma, endometrium carcinoma, chorion carcinoma, pancreaticcarcinoma, prostate carcinoma, testis carcinoma, breast carcinoma,urinary carcinoma, melanoma, brain tumors such as glioblastoma,astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermaltumors, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acutelymphatic leukemia (ALL), chronic lymphatic leukemia (CLL), acutemyeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cellleukemia lymphoma, hepatocellular carcinoma, gall bladder carcinoma,bronchial carcinoma, small cell lung carcinoma, non-small cell lungcarcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma,choroidea melanoma, seminoma, rhabdomyo sarcoma, craniopharyngeoma,osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma,Ewing sarcoma and plasmocytoma.

In particular, EGFR appears to have an important role in the developmentof human brain tumors. A high incidence of overexpression,amplification, deletion and structural rearrangement of the gene codingfor EGFR has been found in biopsies of brain tumors. In fact, theamplification of the EGFR gene in glioblastoma multiforme tumors is oneof the most consistent genetic alterations known, with EGFR beingoverexpressed in approximately 40% of malignant gliomas and EGFRvIIImutation being found in about 50% of all glioblastomas.

In addition to gliomas, abnormal EGFR expression has also been reportedin a number of squamous epidermoid cancers and breast cancers.Interestingly, evidence also suggests that many patients with tumorsthat over-express EGFR have a poorer prognosis than those having tumorsthat do not over-express EGFR.

Non-small cell lung cancer (NSCLC) includes squamous cell carcinomas,adenocarcinoma, bronchioloalveolar carcinoma (BAC), and large cellundifferentiated carcinoma. A subset of patients with NSCLC have beenshown to have mutations in the tyrosine kinase domain of EGFR which isthought to be necessary for the maintenance of the disease. Treatment ofthis subset of patients with NSCLC with gefitinib, a tyrosine kinaseinhibitor which targets EGFR, has shown rapid and dramatic clinicalresponse.

Consequently, therapeutic strategies that can potentially inhibit orreduce the aberrant expression of EGFR are of great interest aspotential anti-cancer agents.

5. Combination Therapies and Treatment of Refractory Cancers

The prophylactic or therapeutic agents of the combination therapies ofthe invention can be administered sequentially or concurrently. In aspecific embodiment, the combination therapies of the invention compriseone or more compounds and at least one other therapy (e.g., anotherprophylactic or therapeutic agent) which has the same mechanism ofaction as said compounds. In another specific embodiment, thecombination therapies of the invention comprise one or more compounds ofthe invention and at least one other therapy (e.g., another prophylacticor therapeutic agent) which has a different mechanism of action thansaid compounds. In certain embodiments, the combination therapies of thepresent invention improve the prophylactic or therapeutic effect of oneor more compounds of the invention by functioning together with thecompounds to have an additive or synergistic effect. In certainembodiments, the combination therapies of the present invention reducethe side effects associated with the therapies (e.g., prophylactic ortherapeutic agents). In certain embodiments, the combination therapiesof the present invention reduce the effective dosage of one or more ofthe therapies.

The prophylactic or therapeutic agents of the combination therapies canbe administered to a subject, preferably a human subject, in the samepharmaceutical composition. In alternative embodiments, the prophylacticor therapeutic agents of the combination therapies can be administeredconcurrently to a subject in separate pharmaceutical compositions. Theprophylactic or therapeutic agents may be administered to a subject bythe same or different routes of administration.

In a specific embodiment, a pharmaceutical composition comprising one ormore compounds of the invention is administered to a subject, preferablya human, to prevent, treat, manage, or ameliorate a proliferativedisorder, such as cancer, or one or more symptom thereof. In accordancewith the invention, pharmaceutical compositions of the invention mayalso comprise one or more other agents (e.g., prophylactic ortherapeutic agents which are currently being used, have been used, orare known to be useful in the prevention, treatment or amelioration of aproliferative disorder or a symptom thereof).

The pharmaceutical compositions can be used in therapy, e.g., to treat amammal with an infection. In one embodiment, the pharmaceuticalcomposition includes one or more additional therapeutic agents, such asone or more additional anti-infective agents.

In another embodiment, the present invention is the use of a compound ofanyone of the formulas disclosed herein for the manufacture of amedicament for treating a mammal with an infection.

In another embodiment of the present invention is a pharmaceuticalcomposition comprising a compound represented by any one of the formulasdisclosed herein and a pharmaceutically acceptable carrier. Thepharmaceutical compositions can be used in therapy, to treat a mammalwith an inflammatory or immune disorder. In one embodiment, thepharmaceutical composition includes one or more additional therapeuticagent, such as one or more additional anti-inflammatory agent or one ormore immunosuppressant.

In, another embodiment, the present invention is the use of a compoundof anyone of the formulas disclosed herein for the manufacture of amedicament for treating a mammal with an inflammatory or autoimmunedisorder or for treatment of a mammal in need of immunosuppression.

The invention provides methods for preventing, managing, treating orameliorating a proliferative disorder, such as cancer, or one or moresymptoms thereof in a subject refractory (either completely orpartially) to existing agent therapies for such a proliferativedisorder, said methods comprising administering to said subject a doseof an effective amount of one or more compounds of the invention and adose of an effective amount of one or more therapies (e.g., one or moreprophylactic or therapeutic agents useful for the prevention, treatment,management, or amelioration of a proliferative disorder or a symptomthereof). The invention also provides methods for preventing, treating,managing, or ameliorating a proliferative disorder or a symptom thereofby administering one or more compounds of the invention in combinationwith any other therapy(ies) to patients who have proven refractory toother therapies but are no longer on these therapies.

The compounds of the invention and/or other therapies can beadministered to a subject by any route known to one of skill in the art.Examples of routes of administration include, but are not limited to,parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g.,inhalation), intranasal, transdermal (topical), transmucosal, and rectaladministration.

6) Agents Useful In Combination With the Compounds of the Invention

Without wishing to be bound by theory, it is believed that the compoundsof the invention can be particularly effective at treating subjectswhose cancer has become multi-drug resistant. Although chemotherapeuticagents initially cause tumor regression, most agents that are currentlyused to treat cancer target only one pathway to tumor progression.Therefore, in many instances, after treatment with one or morechemotherapeutic agents, a tumor develops multidrug resistance and nolonger responds positively to treatment. One of the advantages ofinhibiting Hsp90 activity is that several of its client proteins, whichare mostly protein kinases or transcription factors involved in signaltransduction, have been shown to be involved in the progression ofcancer. Thus, inhibition of Hsp90 provides a method of short circuitingseveral pathways for tumor progression simultaneously. Therefore, it isbelieved that treatment of cancer with an Hsp90 inhibitor of theinvention either alone, or in combination with other chemotherapeuticagents, is more likely to result in regression or elimination of thetumor, and less likely to result in the development of more aggressivemultidrug resistant tumors than other currently available therapies.

In one embodiment, the compounds of the invention can be administeredwith agents that are tyrosine kinase inhibitors (e.g., gefitinib orerlotinib which inhibit EGFR tyrosine kinase activity). In anotherembodiment, the compounds of the invention can be administered topatients whose cancer has become resistant to a tyrosine kinaseinhibitor (e.g., gefitinib or erlotinib). In this embodiment, thecompounds of the invention can be administered either alone or incombination with the tyrosine kinase inhibitor.

In another embodiment, the compounds of the invention are useful fortreating patients with hematological cancers that have become resistantto Imatinib, a chemotherapeutic agent that acts by inhibiting tyrosinekinase activity of Bcr-Abl. In patients with CML in the chronic phase,as well as in a blast crisis, treatment with Imatinib typically willinduce remission. However, in many cases, particularly in those patientswho were in a blast crisis before remission, the remission is notdurable because the Bcr-Abl fusion protein develops mutations in thetyrosine kinase domain that cause it to be resistance to Imatinib. (SeeNimmanapalli, et al., Cancer Research (2001), 61:1799-1804; and Gorre,et al., Blood (2002), 100:3041-3044, the entire teachings of each ofthese references are incorporated herein by reference). Compounds of theinvention act by inhibiting the activity of Hsp90 which disruptBcr-Abl/Hsp90 complexes. When Bcr-Abl is not complex to Hsp90 it israpidly degraded. Therefore, compounds of the invention are effective intreating Imatinib resistant leukemias since they act through a differentmechanism than Imatinib. Compounds of the invention can be administeredalone or with Imatinib in patients who have a Bcr-Abl associated cancerthat is not resistant to Imatinib or to patients whose cancer has becomeresistant to Imatinib.

Anticancer agents that can be co-administered with the compounds of theinvention include Taxol™, also referred to as “paclitaxel”, is awell-known anti-cancer drug which acts by enhancing and stabilizingmicrotubule formation, and analogs of Taxol™, such as Taxotere™.Compounds that have the basic taxane skeleton as a common structurefeature, have also been shown to have the ability to arrest cells in theG2-M phases due to stabilization or inhibition of microtubules.

Other anti-cancer agents that can be employed in combination with thecompounds of the invention include Avastin, Adriamycin, Dactinomycin,Bleomycin, Vinblastine, Cisplatin, acivicin; aclarubicin; acodazolehydrochloride; acronine; adozelesin; aldesleukin; altretamine;ambomycin; ametantrone acetate; aminoglutethimide; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin;dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicinhydrochloride; droloxifene; droloxifene citrate; dromostanolonepropionate; duazomycin; edatrexate; eflornithine hydrochloride;elsamitrucin; enloplatin; enpromate; epipropidine; epirubicinhydrochloride; erbulozole; esorubicin hydrochloride; estramustine;estramustine phosphate sodium; etanidazole; etoposide; etoposidephosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide;floxuridine; fludarabine phosphate; fluorouracil; flurocitabine;fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride;hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmofosine;interleukin II (including recombinant interleukin II, or rIL2),interferon alfa-2a; interferon alfa-2b; interferon alfa-n1; interferonalfa-n3; interferon beta-I a; interferon gamma-I b; iproplatin;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; ormaplatin; oxisuran; pegaspargase;peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman;piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimersodium; porfiromycin; prednimustine; procarbazine hydrochloride;puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide;safingol; safingol hydrochloride; semustine; simtrazene; sparfosatesodium; sparsomycin; spirogermanium hydrochloride; spiromustine;spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin;tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin;teniposide; teroxirone; testolactone; thiamiprine; thioguanine;thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestoloneacetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate;triptorelin; tubulozole hydrochloride; uracil mustard; uredepa;vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate;vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride.

Other anti-cancer drugs that can be employed in combination with thecompounds of the invention include: 20-epi-1,25 dihydroxyvitamin D3;5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine;anagrelide; anastrozole; andrographolide; angiogenesis inhibitors;antagonist D; antagonist G; antarelix; anti-dorsalizing morphogeneticprotein-1; antiandrogen, prostatic carcinoma; antiestrogen;antineoplaston; antisense oligonucleotides; aphidicolin glycinate;apoptosis gene modulators; apoptosis regulators; apurinic acid;ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron;azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat;BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactamderivatives; beta-alethine; betaclamycin B; betulinic acid; bFGFinhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;bistratene A; bizelesin; breflate; bropirimine; budotitane; buthioninesulfoximine; calcipotriol; calphostin C; camptothecin derivatives;canarypox IL-2; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron;doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen;ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur;epirubicin; epristeride; estramustine analogue; estrogen agonists;estrogen antagonists; etanidazole; etoposide phosphate; exemestane;fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;06-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen-binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinasetyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growthinhibitory factor; urokinase receptor antagonists; vapreotide; variolinB; vector system, erythrocyte gene therapy; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer. Preferredanti-cancer drugs are 5-fluorouracil and leucovorin.

Other chemotherapeutic agents that can be employed in combination withthe compounds of the invention include but are not limited to alkylatingagents, antimetabolites, natural products, or hormones. Examples ofalkylating agents useful for the treatment or prevention of T-cellmalignancies in the methods and compositions of the invention includebut are not limited to, nitrogen mustards (e.g., mechloroethamine,cyclophosphamide, chlorambucil, etc.), alkyl sulfonates (e.g.,busulfan), nitrosoureas (e.g., carmustine, lomusitne, etc.), ortriazenes (decarbazine, etc.). Examples of antimetabolites useful forthe treatment or prevention of T-cell malignancies in the methods andcompositions of the invention include but are not limited to folic acidanalog (e.g., methotrexate), or pyrimidine analogs (e.g., Cytarabine),purine analogs (e.g., mercaptopurine, thioguanine, pentostatin).Examples of natural products useful for the treatment or prevention ofT-cell malignancies in the methods and compositions of the inventioninclude but are not limited to vinca alkaloids (e.g., vinblastin,vincristine), epipodophyllotoxins (e.g., etoposide), antibiotics (e.g.,daunorubicin, doxorubicin, bleomycin), enzymes (e.g., L-asparaginase),or biological response modifiers (e.g., interferon alpha).

Examples of alkylating agents that can be employed in combination withthe compounds of the invention include but are not limited to, nitrogenmustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil,melphalan, etc.), ethylenimine and methylmelamines (e.g.,hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan),nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin,etc.), or triazenes (decarbazine, etc.). Examples of antimetabolitesuseful for the treatment or prevention of cancer in the methods andcompositions of the invention include but are not limited to folic acidanalog (e.g., methotrexate), or pyrimidine analogs (e.g., fluorouracil,floxouridine, Cytarabine), purine analogs (e.g., mercaptopurine,thioguanine, pentostatin). Examples of natural-products useful for thetreatment or prevention of cancer in the methods and compositions of theinvention include but are not limited to vinca alkaloids (e.g.,vinblastin, vincristine), epipodophyllotoxins (e.g., etoposide,teniposide), antibiotics (e.g., actinomycin D, daunorubicin,doxorubicin, bleomycin, plicamycin, mitomycin), enzymes (e.g.,L-asparaginase), or biological response modifiers (e.g., interferonalpha). Examples of hormones and antagonists useful for the treatment orprevention of cancer in the methods and compositions of the inventioninclude but are not limited to adrenocorticosteroids (e.g., prednisone),progestins (e.g., hydroxyprogesterone caproate, megestrol acetate,medroxyprogesterone acetate), estrogens (e.g., diethylstilbestrol,ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens (e.g.,testosterone propionate, fluoxymesterone), antiandrogen (e.g.,flutamide), gonadotropin releasing hormone analog (e.g., leuprolide).Other agents that can be used in the methods and compositions of theinvention for the treatment or prevention of cancer include platinumcoordination complexes (e.g., cisplatin, carboblatin), anthracenedione(e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methylhydrazine derivative (e.g., procarbazine), adrenocortical suppressant(e.g., mitotane, aminoglutethimide).

Examples of anti-cancer agents which act by arresting cells in the G2-Mphases due to stabilization or inhibition of microtubules and which canbe used in combination with the compounds of the invention includewithout limitation the following marketed drugs and drugs indevelopment: Erbulozole (also known as R-55104), Dolastatin 10 (alsoknown as DLS-10 and NSC-376128), Mivobulin isethionate (also known asCI-980), Vincristine, NSC-639829, Discodermolide (also known asNVP-XX-A-296), ABT-751 (Abbott, also known as E-7010), Altorhyrtins(such as Altorhyrtin A and Altorhyrtin C), Spongistatins (such asSpongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4,Spongistatin 5, Spongistatin 6, Spongistatin 7, Spongistatin 8; andSpongistatin 9), Cemadotin hydrochloride (also known as LU-103793 andNSC-D-669356), Epothilones (such as Epothilone A, Epothilone B,Epothilone C (also known as desoxyepothilone A or dEpoA), Epothilone D(also referred to as KOS-862, dEpoB, and desoxyepothilone B), EpothiloneE, Epothilone F, Epothilone B N-oxide, Epothilone A N-oxide,16-aza-epothilone B, 21-aminoepothilone B (also known as BMS-310705),21-hydroxyepothilone D (also known as Desoxyepothilone F and dEpoF),26-fluoroepothilone), Auristatin PE (also known as NSC-654663),Soblidotin (also known as TZT-1027), LS-4559-P (Pharmacia, also known asLS-4577), LS-4578 (Pharmacia, also known as LS-477-P), LS-4477(Pharmacia), LS-4559 (Pharmacia), RPR-112378 (Aventis), Vincristinesulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, also known asWS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy ofSciences), BSF-223651 (BASF, also known as ILX-651 and LU-223651),SAH-49960 (Lilly/Novartis), SDZ-268970 (Lilly/Novartis), AM-97(Armad/Kyowa Hakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko),IDN-5005 (Indena), Cryptophycin 52 (also known as LY-355703), AC-7739(Ajinomoto, also known as AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto,also known as AVE-8062, AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A),Vitilevuamide, Tubulysin A, Canadensol, Centaureidin (also known asNSC-106969), T-138067 (Tularik, also known as T-67, TL-138067 andTI-138067), COBRA-1 (Parker Hughes Institute, also known as DDE-261 andWHI-261), H10 (Kansas State University), H16 (Kansas State University),Oncocidin A1 (also known as BTO-956 and DIME), DDE-313 (Parker HughesInstitute), Fijianolide B, Laulimalide, SPA-2 (Parker Hughes Institute),SPA-1 (Parker Hughes Institute, also known as SPIKET-P), 3-IAABU(Cytoskeleton/Mt. Sinai School of Medicine, also known as MF-569),Narcosine (also known as NSC-5366), Nascapine, D-24851 (Asta Medica),A-105972 (Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton/Mt. Sinai Schoolof Medicine, also known as MF-191), TMPN (Arizona State University),Vanadocene acetylacetonate, T-138026 (Tularik), Monsatrol, Inanocine(also known as NSC-698666), 3-IAABE (Cytoskeleton/Mt. Sinai School ofMedicine), A-204197 (Abbott), T-607 (Tularik, also known as T-900607),RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin,Desaetyleleutherobin, Isoeleutherobin A, and Z-Eleutherobin),Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica),D-68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350(Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott),Diozostatin, (−)-Phenylahistin (also known as NSCL-96F037), D-68838(Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris,also known as D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286(also known as SPA-110, trifluoroacetate salt) (Wyeth), D-82317(Zentaris), D-82318 (Zentaris), SC-12983 (NCI), Resverastatin phosphatesodium, BPR-0Y-007 (National Health Research Institutes), and SSR-250411(Sanofi).

7) Anit-Infective Agents Useful In Combination With the Compounds of theInvention

Other anti-fungal agents that can be co-administered with the compoundsof the invention include, but are not limited to, polyene antifungals(e.g., amphotericin and nystatin), azole antifungals (e.g.,ketoconazole, miconazole, fluconazole, itraconazole, posaconazole,ravuconazole, voriconazole, clotrimazole, econazole, oxiconazole,sulconazole, terconazole, butoconazole, and tioconazole), amorolfine,butenafine, naftifine, terbinafine, flucytosine, nikkomycin Z,caspofungin, micafungin (FK463), anidulafungin (LY303366), griseofulvin,ciclopiroxolamine, tolnaftate, intrathecal, haloprogrin, andundecylenate.

Other anti-bacterial agents that can be co-administered with thecompounds of the invention include, but are not limited to, sulfa drugs(e.g., sulfanilamide), folic acid analogs (e.g., trimethoprim),beta-lactams (e.g., penacillin, cephalosporins), aminoglycosides (e.g.,stretomycin, kanamycin, neomycin, gentamycin), tetracyclines (e.g.,chlorotetracycline, oxytetracycline, and doxycycline), macrolides (e.g.,erythromycin, azithromycin, and clarithromycin), lincosamides (e.g.,clindamycin), streptogramins (e.g., quinupristin and dalfopristin),fluoroquinolones (e.g., ciprofloxacin, levofloxacin, and moxifloxacin),polypeptides (e.g., polymixins), rifampin, mupirocin, cycloserine,aminocyclitol (e.g., spectinomycin), glycopeptides (e.g., vancomycin),oxazolidinones (e.g., linezolid), ribosomes, chloramphenicol, fusidicacid, and metronidazole.

Other anti-viral agents that can be co-administered with the compoundsof the invention include, but are not limited to, Emtricitabine (FTC);Lamivudine (3TC); Carbovir; Acyclovir; Interferon; Famciclovir;Penciclovir; Zidovudine (AZT); Didanosine (ddI); Zalcitabine (ddC);Stavudine (d4T); Tenofovir DF (Viread); Abacavir (ABC); L-(−)—FMAU;L-DDA phosphate prodrugs; β-D-dioxolane nucleosides such asβ-D-dioxolanyl-guanine (DG), β-D-dioxolanyl-2,6-diaminopurine (DAPD),and β-D-dioxolanyl-6-chloropurine (ACP); non-nucleoside RT inhibitorssuch as Nevirapine (Viramune), MKC-442, Efavirenz (Sustiva), Delavirdine(Rescriptor); protease inhibitors such as Amprenavir, Atazanavir,Fosamprenavir, Indinavir, Kaletra, Nelfinavir, Ritonavir, Saquinavir,AZT, DMP-450; combination treatments such as Epzicom (ABC+3TC), Trizivir(ABC+3TC+AZT), Truvada (FTC+Viread); Omega IFN (BioMedicines Inc.);BILN-2061 (Boehringer Ingelheim); Summetrel (Endo PharmaceuticalsHoldings Inc.); Roferon A (F. Hoffman-La Roche); Pegasys (F. Hoffman-LaRoche); Pegasys/Ribaravin (F. Hoffman-La Roche); CellCept (F. Hoffman-LaRoche); Wellferon (GlaxoSmithKline); Albuferon-α (Human Genome SciencesInc.); Levovirin (ICN Pharmaceuticals); IDN-6556 (Idun Pharmaceuticals);IP-501 (Indevus Pharmaceuticals); Actimmune (InterMune Inc.); Infergen A(InterMune Inc.); ISIS 14803 (ISIS Pharmaceuticals Inc.); JTK-003 (JapanTobacco Inc.); Pegasys/Ceplene (Maxim Pharmaceuticals); Ceplene (MaximPharmaceuticals); Civacir (Nabi Biopharmaceuticals Inc.); IntronA/Zadaxin (RegeneRx); Levovirin (Ribapharm Inc.); Viramidine (RibapharmInc.); Heptazyme (Ribozyme Pharmaceuticals); Intron A (Schering-Plough);PEG-Intron (Schering-Plough); Rebetron (Schering-Plough); Ribavirin(Schering-Plough); PEG-Intron/Ribavirin (Schering-Plough); Zadazim(SciClone); Rebif (Serono); IFN-β/EMZ701 (Transition Therapeutics); T67(Tularik Inc.); VX-497 (Vertex Pharmaceuticals Inc.); VX-950/LY-570310(Vertex Pharmaceuticals Inc.); Omniferon (Viragen Inc.); XTL-002 (XTLBiopharmaceuticals); SCH 503034 (Schering-Plough); isatoribine and itsprodrugs ANA971 and ANA975 (Anadys); R1479 (Roche Biosciences);Valopicitabine (Idenix); NIM811 (Novartis); Actilon (ColeyPharmaceuticals); Pradefovir (Metabasis Therapeutics); zanamivir;adefovir, adefovir dipivoxil, oseltamivir; vidarabine; gancyclovir;valganciclovir; amantadine; rimantadine; relenza; tamiflu; amantadine;entecavir; and pleconaril.

Other anti-parasitic agents that can be co-administered with thecompounds of the invention include, but are not limited to, avermectins,milbemycins, lufenuron, imidacloprid, organophosphates, pyrethroids,sufanamides, iodquinol, diloxanide furoate, metronidazole, paromycin,azithromycin, quinacrine, furazolidone, tinidazole, ornidazole, bovine,colostrum, bovine dialyzable leukocyte extract, chloroquine, chloroquinephosphate, diclazuril, eflornithine, paromomycin, pentamidine,pyrimethamine, spiramycin, trimethoprim-sulfamethoxazole, albendazole,quinine, quinidine, tetracycline, pyrimethamine-sulfadoxine, mefloquine,doxycycline, proguanil, clindamycin, suramin, melarsoprol, diminazene,nifurtimox, spiroarsoranes, ketoconazole, terbinafine, lovastatin,sodium stibobgluconate, N-methylglucamine antimonate, amphotericin B,allopurinol, itraconazole, sulfadiazine, dapsone, trimetrexate,clarithromycin, roxithromycin, atovaquone, aprinocid, tinidazole,mepacrine hydrochloride, emetine, polyaminopropyl biguanide,paromomycin, benzimidazole, praziquantel, or albendazole.

8) Steroid or Non-Steroidal Anti-Inflammatory Agents Useful inCombination with the Compounds of the Invention

In one embodiment relating to autoimmune, allergic and inflammatoryconditions, the other therapeutic agent may be a steroid or anon-steroidal anti-inflammatory agent. Particularly useful non-steroidalanti-inflammatory agents, include, but are not limited to, aspirin,ibuprofen, diclofenac, naproxen, benoxaprofen, flurbiprofen, fenoprofen,flubufen, ketoprofen, indoprofen, piroprofen, carprofen, oxaprozin,pramoprofen, muroprofen, trioxaprofen, suprofen, aminoprofen,tiaprofenic acid, fluprofen, bucloxic acid, indomethacin, sulindac,tolmetin, zomepirac, tiopinac, zidometacin, acemetacin, fentiazac,clidanac, oxpinac, mefenamic acid, meclofenamic acid, flufenamic acid,niflumic acid, tolfenamic acid, diflurisal, flufenisal, piroxicam,sudoxicam, isoxicam; salicylic acid derivatives, including aspirin,sodium salicylate, choline magnesium trisalicylate, salsalate,diflunisal, salicylsalicylic acid, sulfasalazine, and olsalazin;para-aminophennol derivatives including acetaminophen and phenacetin;indole and indene acetic acids, including indomethacin, sulindac, andetodolac; heteroaryl acetic acids, including tolmetin, diclofenac, andketorolac; anthranilic acids (fenamates), including mefenamic acid, andmeclofenamic acid; enolic acids, including oxicams (piroxicam,tenoxicam), and pyrazolidinediones (phenylbutazone, oxyphenthartazone);and alkanones, including nabumetone and pharmaceutically acceptablesalts thereof and mixtures thereof. For a more detailed description ofthe NSAIDs, see Paul A. Insel, Analgesic-Antipyretic andAntiinflammatory Agents and Drugs Employed in the Treatment of Gout, inGoodman & Gilman's The Pharmacological Basis of Therapeutics 617-57(Perry B. Molinhoff and Raymond W. Ruddon eds., 9^(th) ed 1996) and GlenR. Hanson, Analgesic, Antipyretic and Anti-Inflammatory Drugs inRemington: The Science and Practice of Pharmacy Vol II 1196-1221 (A. R.Gennaro ed. 19th ed. 1995) which are hereby incorporated by reference intheir entireties.

Of particular relevance to allergic disorders, the other therapeuticagent may be an antihistamine. Useful antihistamines include, but arenot limited to, loratadine, cetirizine, fexofenadine, desloratadine,diphenhydramine, chlorpheniramine, chlorcyclizine, pyrilamine,promethazine, terfenadine, doxepin, carbinoxamine, clemastine,tripelennamine, brompheniramine, hydroxyzine, cyclizine, meclizine,cyproheptadine, phenindamine, acrivastine, azelastine, levocabastine,and mixtures thereof. For a more detailed description of antihistamines,see Goodman & Gilman's The Pharmacological Basis of Therapeutics (2001)651-57, 10^(th) ed).

Immunosuppressive agents include glucocorticoids, corticosteroids (suchas Prednisone or Solumedrol), T cell blockers (such as cyclosporin A andFK506), purine analogs (such as azathioprine (Imuran)), pyrimidineanalogs (such as cytosine arabinoside), alkylating agents (such asnitrogen mustard, phenylalanine mustard, buslfan, and cyclophosphamide),folic acid antagonists (such as aminopterin and methotrexate),antibiotics (such as rapamycin, actinomycin D, mitomycin C, puramycin,and chloramphenicol), human IgG, antilymphocyte globulin (ALG), andantibodies (such as anti-CD3 (OKT3), anti-CD4 (OKT4), anti-CD5,anti-CD7, anti-IL-2 receptor, anti-alpha/beta TCR, anti-ICAM-1,anti-CD20 (Rituxan), anti-IL-12 and antibodies to immunotoxins).

E. Compositions and Methods for Administering Therapies

The present invention provides compositions for the treatment,prophylaxis, and amelioration of proliferative disorders, such ascancer. In a specific embodiment, a composition comprises one or morecompounds of the invention, or a pharmaceutically acceptable salt,solvate, clathrate, hydrate or prodrug thereof. In another embodiment, acomposition of the invention comprises one or more prophylactic ortherapeutic agents other than a compound of the invention, or apharmaceutically acceptable salt, solvate, clathrate, hydrate, prodrugthereof. In another embodiment, a composition of the invention comprisesone or more compounds of the invention, or a pharmaceutically acceptablesalt, solvate, clathrate, hydrate or prodrug thereof, and one or moreother prophylactic or therapeutic agents. In another embodiment, thecomposition comprises a compound of the invention, or a pharmaceuticallyacceptable salt, solvate, clathrate, hydrate, or prodrug thereof, and apharmaceutically acceptable carrier, diluent or excipient.

In a preferred embodiment, a composition of the invention is apharmaceutical composition or a single unit dosage form. Pharmaceuticalcompositions and dosage forms of the invention comprise one or moreactive ingredients in relative amounts and formulated in such a way thata given pharmaceutical composition or dosage form can be used to treator prevent proliferative disorders, such as cancer. Preferredpharmaceutical compositions and dosage forms comprise a compound offormula (I)-(VIII), or a pharmaceutically acceptable prodrug, salt,solvate, clathrate, hydrate, or prodrug thereof, optionally incombination with one or more additional active agents.

The pharmaceutical compositions can be used in therapy, e.g., to treat amammal with an infection. In one embodiment, the pharmaceuticalcomposition includes one or more additional therapeutic agents, such asone or more additional anti-infective agents.

In another embodiment, the present invention is the use of a compound ofanyone of the formulas disclosed herein for the manufacture of amedicament for treating a mammal with an infection.

In another embodiment of the present invention is a pharmaceuticalcomposition comprising a compound represented by any one of the formulasdisclosed herein and a pharmaceutically acceptable carrier. Thepharmaceutical compositions can be used in therapy, e.g., to treat amammal with an inflammatory or immune disorder. In one embodiment, thepharmaceutical composition includes one or more additional therapeuticagent, such as one or more additional anti-inflammatory agent or one ormore immunosuppressant.

In another embodiment, the present invention is the use of a compound ofanyone of the formulas disclosed herein for the manufacture of amedicament for treating a mammal with an inflammatory or autoimmunedisorder or for treatment of a mammal in need of immunosuppression.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include, but are not limited to, parenteral, e.g.,intravenous, intradermal, subcutaneous, oral (e.g., inhalation),intranasal, transdermal (topical), transmucosal, and rectaladministration. In a specific embodiment, the composition is formulatedin accordance with routine procedures as a pharmaceutical compositionadapted for intravenous, subcutaneous, intramuscular, oral, intranasalor topical administration to human beings. In a preferred embodiment, apharmaceutical composition is formulated in accordance with routineprocedures for subcutaneous administration to human beings.

Single unit dosage forms of the invention are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), or transdermal administration to a patient. Examples ofdosage forms include, but are not limited to: tablets; caplets;capsules, such as soft elastic gelatin capsules; cachets; troches;lozenges; dispersions; suppositories; ointments; cataplasms (poultices);pastes; powders; dressings; creams; plasters; solutions; patches;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; and sterile solids (e.g., crystalline or amorphous solids)that can be reconstituted to provide liquid dosage forms suitable forparenteral administration to a patient.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage formsuitable for mucosal administration may contain a smaller amount ofactive ingredient(s) than an oral dosage form used to treat the sameindication. This aspect of the invention will be readily apparent tothose skilled in the art. See, e.g., Remington's Pharmaceutical Sciences(1990) 18th ed., Mack Publishing, Easton Pa.

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsare provided herein. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a patient.For example, oral dosage forms such as tablets may contain excipientsnot suited for use in parenteral dosage forms.

The suitability of a particular excipient may also depend on thespecific active ingredients in the dosage form. For example, thedecomposition of some active ingredients can be accelerated by someexcipients such as lactose, or when exposed to water. Active ingredientsthat comprise primary or secondary amines (e.g., N-desmethylvenlafaxineand N,N-didesmethylvenlafaxine) are particularly susceptible to suchaccelerated decomposition. Consequently, this invention encompassespharmaceutical compositions and dosage forms that contain little, ifany, lactose. As used herein, the term “lactose-free” means that theamount of lactose present, if any, is insufficient to substantiallyincrease the degradation rate of an active ingredient. Lactose-freecompositions of the invention can comprise excipients that are wellknown in the art and are listed, for example, in the U.S. Pharmocopia(USP)SP(XXI)/NF (XVI). In general, lactose-free compositions compriseactive ingredients, a binder/filler, and a lubricant in pharmaceuticallycompatible and pharmaceutically acceptable amounts. Preferredlactose-free dosage forms comprise active ingredients, microcrystallinecellulose, pre-gelatinized starch, and magnesium stearate.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen (1995) Drug Stability: Principles &Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 379-80. In effect, water andheat accelerate the decomposition of some compounds. Thus, the effect ofwater on a formulation can be of great significance since moistureand/or humidity are commonly encountered during manufacture, handling,packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are preferablyanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical Composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizer” include, but are not limited to, antioxidantssuch as ascorbic acid, pH buffers, or salt buffers.

1). Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such, as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences (1990) 18th ed., MackPublishing, Easton Pa.

Typical oral dosage forms of the invention are prepared by combining theactive ingredient(s) in an admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration. For example, excipients suitablefor use in oral liquid or aerosol dosage forms include, but are notlimited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Onespecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103J and Starch 1500LM.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form. Disintegrants are used in thecompositions of the invention to provide tablets that disintegrate whenexposed to an aqueous environment. Tablets that contain too muchdisintegrant may disintegrate in storage, while those that contain toolittle may not disintegrate at a desired rate or under the desiredconditions. Thus, a sufficient amount of disintegrant that is neithertoo much nor too little to detrimentally alter the release of the activeingredients should be used to form solid oral dosage forms of theinvention. The amount of disintegrant used varies based upon the type offormulation, and is readily discernible to those of ordinary skill inthe art. Typical pharmaceutical compositions comprise from about 0.5 toabout 15 weight percent of disintegrant, preferably from about 1 toabout 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, other starches, pre-gelatinizedstarch, other starches, clays, other algins, other celluloses, gums, andmixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

2) Controlled Release Dosage Forms

Active ingredients of the invention can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

A particular extended release formulation of this invention comprises atherapeutically or prophylactically effective amount of a compound offormula (I)-(VIII), or a pharmaceutically acceptable salt, solvate,hydrate, clathrate, or prodrug thereof, in spheroids which furthercomprise microcrystalline cellulose and, optionally,hydroxypropylmethyl-cellulose coated with a mixture of ethyl celluloseand hydroxypropylmethylcellulose. Such extended release formulations canbe prepared according to U.S. Pat. No. 6,274,171, the entirely of whichis incorporated herein by reference.

A specific controlled-release formulation of this invention comprisesfrom about 6% to about 40% a compound of formula (I)-(VIII), or apharmaceutically acceptable salt, solvate, hydrate, clathrate, orprodrug thereof, by weight, about 50% to about 94% microcrystallinecellulose, NF, by weight, and optionally from about 0.25% to about 1% byweight of hydroxypropyl-methylcellulose, USP, wherein the spheroids arecoated with a film coating composition comprised of ethyl cellulose andhydroxypropylmethylcellulose.

3) Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP;aqueous-vehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention.

4) Transdermal, Topical, and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms of the invention include,but are not limited to, ophthalmic solutions, sprays, aerosols, creams,lotions, ointments, gels, solutions, emulsions, suspensions, or otherforms known to one of skill in the art. See, e.g., Remington'sPharmaceutical Sciences (1980 & 1990) 16th and 18th eds., MackPublishing, Easton Pa. and Introduction to Pharmaceutical Dosage Forms(1985) 4th ed., Lea & Febiger, Philadelphia. Dosage forms suitable fortreating mucosal tissues within the oral cavity can be formulated asmouthwashes or as oral gels. Further, transdermal dosage forms include“reservoir type” or “matrix type” patches, which can be applied to theskin and worn for a specific period of time to permit the penetration ofa desired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms encompassed by this invention are well known to those skilled inthe pharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof to form lotions, tinctures, creams, emulsions, gelsor ointments, which are non-toxic and pharmaceutically acceptable.Moisturizers or humectants can also be added to pharmaceuticalcompositions and dosage forms if desired. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences (1980 & 1990) 16th and 18th eds., MackPublishing, Easton Pa.

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, oleyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

5) Dosage & Frequency of Administration

The amount of the compound or composition of the invention which will beeffective in the prevention, treatment, management, or amelioration of aproliferative disorders, such as cancer, or one or more symptomsthereof, will vary with the nature and severity of the disease orcondition, and the route by which the active ingredient is administered.The frequency and dosage will also vary according to factors specificfor each patient depending on the specific therapy (e.g., therapeutic orprophylactic agents) administered, the severity of the disorder,disease, or condition, the route of administration, as well as age,body, weight, response, and the past medical history of the patient.Effective doses may be extrapolated from dose-response curves derivedfrom in vitro or animal model test systems. Suitable regiments can beselected by one skilled in the art by considering such factors and byfollowing, for example, dosages reported in the literature andrecommended in the Physician's Desk Reference (57th ed., 2003).

Exemplary doses of a small molecule include milligram or microgramamounts of the small molecule per kilogram of subject or sample weight(e.g., about 1 microgram per kilogram to about 500 milligrams perkilogram, about 100 micrograms per kilogram to about 5 milligrams perkilogram, or about 1 microgram per kilogram to about 50 micrograms perkilogram).

In general, the recommended daily dose range of a compound of theinvention for the conditions described herein lie within the range offrom about 0.01 mg to about 1000 mg per day, given as a singleonce-a-day dose preferably as divided doses throughout a day. In oneembodiment, the daily dose is administered twice daily in equallydivided doses. Specifically, a daily dose range should be from about 5mg to about 500 mg per day, more specifically, between about 10 mg andabout 200 mg per day. In managing the patient, the therapy should beinitiated at a lower dose, perhaps about 1 mg to about 25 mg, andincreased if necessary up to about 200 mg to about 1000 mg per day aseither a single dose or divided doses, depending on the patient's globalresponse. It may be necessary to use dosages of the active ingredientoutside the ranges disclosed herein in some cases, as will be apparentto those of ordinary skill in the art. Furthermore, it is noted that theclinician or treating physician will know how and when to interrupt,adjust, or terminate therapy in conjunction with individual patientresponse.

Different therapeutically effective amounts may be applicable fordifferent proliferative disorders, as will be readily known by those ofordinary skill in the art. Similarly, amounts sufficient to prevent,manage, treat or ameliorate such proliferative disorders, butinsufficient to cause, or sufficient to reduce, adverse effectsassociated with the compounds of the invention are also encompassed bythe above described dosage amounts and dose frequency schedules.Further, when a patient is administered multiple dosages of a compoundof the invention, not all of the dosages need be the same. For example,the dosage administered to the patient may be increased to improve theprophylactic or therapeutic effect of the compound or it may bedecreased to reduce one or more side effects that a particular patientis experiencing.

In a specific embodiment, the dosage of the composition of the inventionor a compound of the invention administered to prevent, treat, manage,or ameliorate a proliferative disorders, such as cancer, or one or moresymptoms thereof in a patient is 150 μg/kg, preferably 250 μg/kg, 500μg/kg, 1 mg/kg, 5 mg/kg, 10 mg/kg, 25 mg/kg, 50 mg/kg, 75 mg/kg, 100mg/kg, 125 mg/kg, 150 mg/kg, or 200 mg/kg or more of a patient's bodyweight. In another embodiment, the dosage of the composition of theinvention or a compound of the invention administered to prevent, treat,manage, or ameliorate a proliferative disorders, such as cancer, or oneor more symptoms thereof in a patient is a unit dose of 0.1 mg to 20 mg,0.1 mg to 15 mg, 0.1 mg to 12 mg, 0.1 mg to 10 mg, 0.1 mg to 8 mg, 0.1mg to 7 mg, 0.1 mg to 5 mg, 0.1 to 2.5 mg, 0.25 mg to 20 mg, 0.25 to 15mg, 0.25 to 12 mg, 0.25 to 10 mg, 0.25 to 8 mg, 0.25 mg to 7 mg, 0.25 mgto 5 mg, 0.5 mg to 2.5 mg, 1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 12 mg,1 mg to 10 mg, 1 mg to 8 mg, 1 mg to 7 mg, 1 mg to 5 mg, or 1 mg to 2.5mg.

The dosages of prophylactic or therapeutic agents other than compoundsof the invention, which have been or are currently being used toprevent, treat, manage, or proliferative disorders, such as cancer, orone or more symptoms thereof can be used in the combination therapies ofthe invention. Preferably, dosages lower than those which have been orare currently being used to prevent, treat, manage, or ameliorate aproliferative disorders, or one or more symptoms thereof, are used inthe combination therapies of the invention. The recommended dosages ofagents currently used for the prevention, treatment, management, oramelioration of a proliferative disorders, such as cancer, or one ormore symptoms thereof, can obtained from any reference in the artincluding, but not limited to, Hardman et al., eds., 1996, Goodman &Gilman's The Pharmacological Basis Of Basis Of Therapeutics 9^(th) Ed,Mc-Graw-Hill, New York; Physician's Desk Reference (PDR) 57^(th) Ed.,2003, Medical Economics Co., Inc., Montvale, N.J., which areincorporated herein by reference in its entirety.

In certain embodiments, when the compounds of the invention areadministered in combination with another therapy, the therapies (e.g.,prophylactic or therapeutic agents) are administered less than 5 minutesapart, less than 30 minutes apart, 1 hour apart, at about 1 hour apart,at about 1 to about 2 hours apart, at about 2 hours to about 3 hoursapart, at about 3 hours to about 4 hours apart, at about 4 hours toabout 5 hours apart, at about 5 hours to about 6 hours apart, at about 6hours to about 7 hours apart, at about 7 hours to about 8 hours apart,at about 8 hours to about 9 hours apart; at about 9 hours to about 10hours apart, at about 10 hours to about 11 hours apart, at about 11hours to about 12 hours apart, at about 12 hours to 18 hours apart, 18hours to 24 hours apart, 24 hours to 36 hours apart, 36 hours to 48hours apart, 48 hours to 52 hours apart, 52 hours to 60 hours apart, 60hours to 72 hours apart, 72 hours to 84 hours apart, 84 hours to 96hours apart, or 96 hours to 120 hours part. In one embodiment, two ormore therapies (e.g., prophylactic or therapeutic agents) areadministered within the same patent visit.

In certain embodiments, one or more compounds of the invention and oneor more other the therapies (e.g., prophylactic or therapeutic agents)are cyclically administered. Cycling therapy involves the administrationof a first therapy (e.g., a first prophylactic or therapeutic agents)for a period of time, followed by the administration of a second therapy(e.g., a second prophylactic or therapeutic agents) for a period oftime, followed by the administration of a third therapy (e.g., a thirdprophylactic or therapeutic agents) for a period of time and so forth,and repeating this sequential administration, i.e., the cycle in orderto reduce the development of resistance to one of the agents, to avoidor reduce the side effects of one of the agents, and/or to improve theefficacy of the treatment.

In certain embodiments, administration of the same compound of theinvention may be repeated and the administrations may be separated by atleast 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days,2 months, 75 days, 3 months, or 6 months. In other embodiments,administration of the same prophylactic or therapeutic agent may berepeated and the administration may be separated by at least at least 1day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2months, 75 days, 3 months, or 6 months.

In a specific embodiment, the invention provides a method of preventing,treating, managing, or ameliorating a proliferative disorders, such ascancer, or one or more symptoms thereof, said methods comprisingadministering to a subject in need thereof a dose of at least 150 μg/kg,preferably at least 250 μg/kg, at least 500 μg/kg, at least 1 mg/kg, atleast 5 mg/kg, at least 10 mg/kg, at least 25 mg/kg, at least 50 mg/kg,at least 75 mg/kg, at least 100 mg/kg, at least 125 mg/kg, at least 150mg/kg, or at least 200 mg/kg or more of one or more compounds of theinvention once every day, preferably, once every 2 days, once every 3days, once every 4 days, once every 5 days, once every 6 days, onceevery 7 days, once every 8 days, once every 10 days, once every twoweeks, once every three weeks, or once a month.

F. Other Embodiments

The compounds of the invention may be used as research tools (forexample, to evaluate the mechanism of action of new drug agents, toisolate new drug discovery targets using affinity chromatography, asantigens in an ELISA or ELISA-like assay, or as standards in in vitro orin vivo assays). These and other uses and embodiments of the compoundsand compositions of this invention will be apparent to those of ordinaryskill in the art.

The invention is further defined by reference to the following examplesdescribing in detail the preparation of compounds of the invention. Itwill be apparent to those skilled in the art that many modifications,both to materials and methods, may be practiced without departing fromthe purpose and interest of this invention. The following examples areset forth to assist in understanding the invention and should not beconstrued as specifically limiting the invention described and claimedherein. Such variations of the invention, including the substitution ofall equivalents now known or later developed, which would be within thepurview of those skilled in the art, and changes in formulation or minorchanges in experimental design, are to be considered to fall within thescope of the invention incorporated herein.

EXAMPLES Example 14-(4-(2,3-dihydro-1H-inden-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyldihydrogen phosphate (Compound 1)

The synthesis consists of 5 steps. The following is the procedure:

Synthesis of the thioamideN-(2,3-dihydro-1H-inden-5-yl)-2,4-dihydroxy-5-isopropylbenzothioamide 3

To a stirred suspension of 0.50 g (3.28 mmols) of 4-isopropyl resorcinolin 5 mL of BF₃.2AcOH at 5° C. was added drop wise 0.57 g (3.28 mmols) of5-isothiocyanato-2,3-dihydro-1H-indene 2. The mixture was stirred at 5°C. for 15 min. and cooling bath removed. The mixture was then stirred atroom temperature for 1.5 h, poured into 100 mL of cold water whilestirring. The yellow solid obtained was filtered, dried, redissolved inethylacetate and dried over Na₂SO₄. Concentration followed by columnchromatography afforded 0.70 g of the product 3 as yellow solid.

Synthesis of3-(2,3-dihydro-1H-inden-5-yl)-7-hydroxy-6-isopropyl-4-thioxo-3,4-dihydro-2H-benzo[e][1,3]oxazin-2-one5

To a stirred solution of 0.20 g (0.61 mmols) of the thioamide 3 in 6 mLof anhydrous tetrahydrofuran at room temperature, was added 99 mg (0.61mmols) of carbonyldiimidazole 4 portion wise. The resultant mixture wasstirred at room temperature for 2 h and concentrated. Filtration of theproduct through a short pad of silica gel eluting with 3:1hexane:ethylacetate afforded 0.23 g of the product 5 as bright yellowsolid.

Synthesis of dibenzyl3-(2,3-dihydro-1,1-inden-5-yl)-6-isopropyl-2-oxo-4-thioxo-3,4-dihydro-2H-benzo[e][1,3]oxazin-7-ylphosphate 7

To a stirred solution of 0.30 g (0.84 mmols) of 5 in 10 mL of anhydrousacetonitrile at 5° C., was added 0.4 mL (4.24 mmols) of CCl₄, 0.31 mL(1.78 mmols) of diisopropylethylamine and 10 mg (85□mols) of4-dimethylaminopyridine in that order. After stirring the mixture for 5min. at 5° C., 0.34 mL (1.27 mmols) of dibenzyl phosphonate 6 was addeddrop wise and the mixture stirred for 1 h at 5° C. The reaction wasquenched with water, extracted the product with ethyl acetate andorganic layers were washed with water and brine. Filtration of theproduct through a plug of silica gel afforded 0.52 g of the product 7 asbrown liquid.

Synthesis of dibenzyl4-(4-(2,3-dihydro-1H-inden-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenylphosphate 8

To a stirred solution of 0.52 g (0.84 mmols) of 7 in 15 mL of anhydrousethanol was added 63 mg (1.27 mmols) of hydrazine hydrate and themixture stirred at 80° C. for 30 min. The mixture was concentrated andchromatographed on silica gel using 1:1 hexane:ethylacetate to afford0.16 g of the product 8 as off-white foam.

Synthesis of4-(4-(2,3-dihydro-1H-inden-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyldihydrogen phosphate 9

A solution of 0.15 g (0.24 mmols) of 8 and 25 mg of Pd—C catalyst in 20mL of anhydrous ethanol was subjected to hydrogenation conditions at 1atm pressure for 2 h. The catalyst was filtered off and the crudeproduct was reslurried in 98:2 mixture of diethylether:ethanol to afford90 mg of the phosphate 9 as off white solid.

¹H NMR (300 MHz, DMSO-d₆) δ 11.95 (s, 1H), 9.76 (s, 1H), 7.16 (d, J=7.8Hz, 1H), 7.08 (s, 1H) 7.02 (s, 1H), 6.86 (d, J=7.5 Hz, 1H), 6.79 (s,1H), 3.14-3.04 (m, 1H), 2.84-2.77 (m, 4H), 2.04-1.94 (m, 2H), 1.01 (d,J=6.6 Hz, 6H)

ESMS calcd for C20H22N₃O6P: 431.12; Found: 432.2 (M+1)

Example 25-hydroxy-4-(5-hydroxy-4-(6-morpholinopyridin-3-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyldihydrogen phosphate (Compound 2)

6-morpholinopyridin-3-amine (3.59 g, 20 mmol) in 50 ml CH₂Cl₂ was addedto the solution N,N′-carbonyldiimidazole (CDI) (4.0 g, 25 mmol) in 100ml CH₂Cl₂, and the solution was mix at RT for 1 h. Solvent were removedon rotary evaporator and the residue was dissolved in 50 ml dioxane andtreated with hydrazine (6.5 ml, 200 mmol, 10 eq) at RT to 45° C. for 1h. The reaction mixture was subjected to EtOAc/aqueous workup to removeexcess hydrazine The aqueous solution was extracted with CH₂Cl₂ Theorganic layer was dried over Na₂SO₄, filtered evaporated in vacuo gaveN-(6-morpholinopyridin-3-yl)hydrazinecarboxamide (1) (3.32 g, 70%) as alight brown solid. To a stirred solution of 1.90 g (6.0 mmol) of the2-(benzyloxy)-5-isopropyl-4-(methoxymethoxy)benzaldehyde (2) in 40 ml ofmethanol was added 1.43 g (6.0 mmol) of the hydrazide (1) and aceticacid (3 drop). The resultant mixture was then heated at 60° C. for 1 h,then cooled. The white precipitate thus obtained was filtered, washedwith ether (2×20 ml) gave 2.88 g (90%) of(E)-2-(2-(benzyloxy)-5-isopropyl-4-(methoxymethoxy)benzylidene)-N-(6-morpholinopyridin-3-yl)hydrazinecarboxamide(3) as white solid.

To a solution of 2.40 g (4.5 mmol) of benzylidene-carboxamide (3) and0.55 g (13.75 mmol) of NaOH in 75 ml of EtOH, was added 3.3 g (10.0mmol) of K₃Fe(CN)₆ at once. The resultant mixture was then refluxed for8 h and the inorganics were filtered off. Concentration of the filtrate,addition of 35 mL of water and acidification using 2N HCl till pH 7-8afforded pale brown precipitate. It was filtered, washed with water anddried. The resultant crude solid was then purification by silica gelchromatography (elution with 1:4 and 1:1 ethyl acetate/hexane and ethylacetate) gave 2.14 g, (89%).5-(2-(benzyloxy)-5-isopropyl-4-(methoxymethoxy)phenyl)-4-(6-morpholinopyridin-3-yl)-4H-1,2,4-triazol-3-ol(4) as white solid.

To the solution of compound (4) (2.10 g, 3.95 mmol) in MeOH (100 ml),HCl (2 ml, 2N) was added, and then the resultant mixture was heated at65° C. for 5 h, then cooled. The white precipitate thus obtained wasfiltered, washed with ether (2×20 ml) gave 1.53 g (74%) of5-(2-(benzyloxy)-4-hydroxy-5-isopropylphenyl)-4-(6-morpholinopyridin-3-yl)-4H-1,2,4-triazol-3-olhydrochloride (5).

To the solution of compound (5) (1.53 g, 2.9 mmol) in CH₃CN (25 ml),CCl₄ (2.41 ml, 25 mmol) was added, and then cooled to 0° C. To thatmixture first N,N-diisopropylethylamine (3.5 ml, 20 mmol), andN,N-dimethylaminopyridine (85 mg, 0.7 mmol) was added, and one minutelater dibenzyl phosphite (1.84 g, 7.0 mmol) were dropwise addedsuccessively. The reaction mixture was then stirred at 0° C. for 1 hr.When the reaction has been completed (monitored by LC/MS) water (40 ml)was added and the mixture was extracted by ethyl acetate (3×40 ml). Thecombined organic phase was washed with saturated aq_.NaCl, dried overNa₂SO₄ and concentrated in vacuo. The crude mixture was purified byflash chromatography (hexane-EtOAc 3:1) to give 2.3 g of mixturedibenzyl5-(benzyloxy)-4-(5-hydroxy-4-(6-morpholinopyridin-3-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenylphosphate (6) whit small amount dibenzyl5-(benzyloxy)-4-(1-(bis(benzyloxy)phosphoryl)-4-(6-morpholinopyridin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-isopropylphenylphosphate.

Mixture (6) (2.3 g) was hydrogenated in MeOH (50 mL) using Pd/C (10%,dry, 200 mg) and H₂ balloon at 1 atm at room temperature for 2 hr. Pd/Cwas filtered off through a pad of celite and the mother liquid wasconcentrated to give Compound 2 as white solid (1.13 g, 81%).

¹H-NMR (DMSO-d₆) δ (ppm), 11.89 (s, 1H), 9.60 (s, 1H), 7.87 (s, 1H),7.35 (d, J=2.7, J=9.0 Hz, 1H)), 6.92 (s, 1H), 6.81 (d, J=9.0 Hz, 1H),6.24 (s, 1H), 3.67-3.64 (m, 4H), 3.43-3.40 (m, 4H), 3.08-2.95 (m, 1H),1.04 (d, J=6:9 Hz, 6H);

ESMS clcd for C₂₀H₂₄N₅O₇P: 477.14; Found: 478.2 (M+1)⁺.

Example 35-hydroxy-2-isopropyl-4-(4-(1-methyl-1H-indol-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate (Compound 3)

(E)-2-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylbenzylidene)-N-(1-methyl-1H-indol-5-yl)hydrazinecarboxamide

To a suspension of the aldehyde in ethanol was added AcOH and stirred.To the resultant mixture was added ofN-(1-methyl-1H-indol-5-yl)hydrazinecarboxamide portion wise at roomtemperature and the resultant mixture was heated at 80° C. for 1 h. Themixture was cooled to RT and filtered the precipitate, washed withethanol and ether and dried. Vacuum drying afforded the product.

3-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylphenyl)-4-(1-methyl-1H-indol-5-yl)-1H-1,2,4-triazol-5(4H)-one

To a stirred suspension of(E)-2-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylbenzylidene)-N-(1-methyl-1H-indol-5-yl)hydrazinecarboxamidein ethanol was added NaOH and stirred. To the resultant mixture, wasadded K₃Fe(CN)₆ at once and the resultant mixture was stirred at refluxtemperature till the reaction is complete, checked by TLC. The mixturewas cooled and the inorganics were filtered off. The residues werethoroughly washed with EtOH and filtrates were collected. The combinedfiltrates were concentrated and crude mixture was washed with water andextracted with EtOAc. Drying and concentration produced brown solid,which was filtered and washed with ether. The crude was carried out tothe next reaction without further purification.

benzyl3-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylphenyl)-4-(1-methyl-1H-indol-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1-carboxylate

Cbz-Cl was added to the solution of3-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylphenyl)-4-(1-methyl-1H-indol-5-yl)-1H-1,2,4-triazol-5(4H)-oneand DIPEA in dichloromethane and reaction solution was stirred for 2 hat room temperature. Washed with water and extracted withdichloromethane and solvent was removed. The crude reaction mixture wascarried over to the next reaction without further purification. Thereaction mixture was diluted by methanol and treated with concentratedHCl and heated it up to 65° C. Column chromatogram produced the desiredbenzyl3-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylphenyl)-4-(1-methyl-1H-indol-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1-carboxylate.

5-hydroxy-2-isopropyl-4-(4-(1-methyl-1H-indol-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate

Dibenzyl phosphorochloridate was added to the solution of benzyl3-(2-(benzyloxy)-4-(ethoxymethoxy)-5-isopropylphenyl)-4-(1-methyl-1H-indol-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1-carboxylateand potassium carbonate in acetone. The reaction mixture was stirred atroom temperature overnight. After removal of solvent, the reactionmixture was purified by column chromatography to produce pale yellowoil, which was hydrogenated to give the desired.

Example A Inhibition of Hsp90

Hsp90 protein is obtained from Stressgen (Cat#SPP-770). Assay buffer:100 mM Tris-HCl, Ph7.4, 20 mM KCl, 6 mM MgCl₂. Malachite green (0.0812%w/v) (M9636) and polyviny alcohol USP (2.32% w/v) (P1097) are obtainedfrom Sigma. A Malachite Green Assay (see Methods Mol Med, 2003, 85:149for method details) is used for examination of ATPase activity of Hsp90protein. Briefly, Hsp90 protein in assay buffer (100 mM Tris-HCl, Ph7.4,20 mM KCl, 6 mM MgCl₂) is mixed with ATP alone (negative control) or inthe presence of Geldanamycin (a positive control) or a compound of theinvention in a 96-well plate. Malachite green reagent is added to thereaction. The mixtures are incubated at 37° C. for 4 hours and sodiumcitrate buffer (34% w/v sodium citrate) is added to the reaction. Theplate is read by an ELISA reader with an absorbance at 620 nm.

Example B Degradation of Hsp90 Client Proteins via Inhibition of Hsp90Activity A. Cells and Cell Culture

Human high-Her2 breast carcinoma BT474 (HTB-20), SK-BR-3 (HTB-30) andMCF-7 breast carcinoma (HTB-22) from American Type Culture Collection,VA, USA were grown in Dulbecco's Modified Eagle's medium with 4 mML-glutamine and antibiotics (100 IU/ml penicillin and 100 ug/mlstreptomycine;GibcoBRL). To obtain exponential cell growth, cells weretrypsinized, counted and seeded at a cell density of 0.5×10⁶ cells/mlregularly, every 3 days. All experiments were performed on day 1 aftercell passage.

B. Degradation of Her2 in Cells after Treatment with a Compound of theInvention

1. Method 1

BT-474 cells were treated with 0.5 μM, 2 μM, or 5 μM of 17AAG (apositive control) or 0.5 μM, 2 μM, or 5 μM of a compound of theinvention overnight in DMEM medium. After treatment, each cytoplasmicsample was prepared from 1×10⁶ cells by incubation of cell lysis buffer(#9803, cell Signaling Technology) on ice for 10 minutes. The resultingsupernatant used as the cytosol fractions are dissolved with samplebuffer for SDS-PAGE and run on a SDS-PAGE gel, blotted onto anitrocellulose membrane by using semi-dry transfer. Non-specific bindingto nitrocellulose was blocked with 5% skim milk in TBS with 0.5% Tweenat room temperature for 1 hour, then probed with anti-Her2/ErB2 mAb(rabbit IgG, #2242, Cell Signaling) and anti-Tubulin (T9026, Sigma) ashousekeeping control protein. HRP-conjugated goat anti-rabbit IgG (H+L)and HRP-conjugated horse anti-mouse IgG (H+L) were used as secondary Ab(#7074, #7076, Cell Signaling) and LumiGLO reagent, 20× Peroxide (#7003,Cell Signaling) was used for visualization. The results obtained byemploying compounds 1 through 4 are presented in Table 1.

TABLE 1 Compound IC50 Data (nM) 1 228 2 90 3 88 4 182

As seen from Table 1, Her2, an Hsp90 client protein, is degraded whencells are treated with compounds of the invention. 0.5 μM of 17AAG, aknown Hsp90 inhibitor which is used as a positive control, causespartial degradation of Her2 (results not shown).

2. Method 2

MV-4-11 cells (20,000 cells/well) are cultured in 96-well plates andmaintained at 37° C. for several hours. The cells are treated with acompound of the invention or 17AAG (a positive control) at variousconcentrations and incubated at 37° C. for 72 hours. Cell survival ismeasured with Cell Counting Kit-8 (Dojindo Laboratories, Cat. # CK0

Fluorescent Staining of Her2 on the Surface of Cells Treated with aCompound of the Invention

After treatment with a compound of the invention, cells are washed twicewith 1×PBS/1% FBS, and then stained with anti-Her2-FITC (#340553, BD)for 30 min at 4° C. Cells are then washed three times in FACS bufferbefore the fixation in 0.5 ml 1% paraformadehydrede. Data is acquired ona FACSCalibur system. Isotype-matched controls are used to establish thenon-specific staining of samples and to set the fluorescent markers. Atotal 10,000 events were recorded from each sample. Data are analysed byusing CellQuest software (BD Biosciences).

D. Apoptosis Analysis

After treatment with the compounds of the invention, cells are washedonce with 1×PBS/1% FBS, and then stained in binding buffer withFITC-conjugated Annexin V and Propidium iodide (PI) (all obtained fromBD Biosciences) for 30 min at 4° C. Flow cytometric analysis isperformed with FACSCalibur (BD Biosciences) and a total 10,000 eventsare recorded from each sample. Data is analyzed by using CellQuestsoftware (BD Biosciences). The relative fluorescence is calculated aftersubtraction of the fluorescence of control.

E. Degradation of c-Kit in Cells after Treatment with a Compound of theInvention

Two leukemia cell lines, HEL92.7.1 and Kasumi-1, are used for testingc-kit degradation induced by Hsp90 inhibitors of the invention. Thecells (3×10⁵ per well) are treated with 17AAG (0.5 μM), or a compound ofthe invention for about 18 h. The cells are collected and centrifuged(SORVALL RT 6000D) at 1200 rpm for 5 min. The supernatants arediscarded, and the cells are washed one time with 1×PBS. Aftercentrifugation the cells are stained with FITC conjugated c-kit antibody(MBL International, Cat# K0105-4) in 100 ml 1×PBS at 4° C. for 1 h. Thesamples are read and analysized with FACSCalibur flow cytometer (BectonDicknson).

c-Kit, a tyrosine kinase receptor and one of the Hsp90 client proteins,is selected and used in a FACS-based degradation assay. Compounds of theinvention are expected to induce c-kit degradation in a dose-dependentmanner. Compounds of the invention are expected to be effective in thetreatment of c-kit associated tumors, such as leukemias, mast celltumors, small cell lung cancer, testicular cancer, some cancers of thegastrointestinal tract (including GIST), and some central nervoussystem.

The results of the FACS analysis can be confirmed with Western blotanalysis.

F. Degradation of c-Met in Cells after Treatment with a Compound of theInvention

The ability of the Hsp90 inhibitors of the invention to induce thedegradation of c-Met, an Hsp90 client protein that is expressed at highlevels in several types of non-small cell lung cancer can be examined.NCI-H1993 (ATCC, cat# CRL-5909) are seeded in 6-well plates at 5×10⁵cells/well. The cells are treated with 17AAG (100 nM or 400 nM) or acompound of the invention (100 nM or 400 nM), and cell lysis is prepared24 h after treatment. Equal amount of proteins are used for Western blotanalysis. The compounds of the invention are expected to potently inducedegradation of c-Met in this cell line due to inhibition of Hsp90.

Example C Anti-tumor Activity Against the Human Tumor Cell LineMDA-MB-435S in a nude Mouse Xenograft Model

The human tumor cell line, MDA-MB-435S (ATCC #HTB-129; G. Ellison, etal., Mol. Pathol. 55:294-299, 2002), is obtained from the American TypeCulture Collection (Manassus, Va., USA). The cell line is cultured ingrowth media prepared from 50% Dulbecco's Modified Eagle Medium (highglucose), 50% RPMI Media 1640, 10% fetal bovine serum (FBS), 1%100×L-glutamine, 1% 100× Penicillin-Streptomycin, 1% 100× sodiumpyruvate and 1% 100×MEM non-essential amino acids. FBS is obtained fromSigma-Aldrich Corp. (St. Louis, Mo., USA), and all other reagents areobtained from Invitrogen Corp. (Carlsbad, Calif., USA). Approximately4-5×10(6) cells that have been cryopreserved in liquid nitrogen arerapidly thawed at 37° C. and transferred to a 175 cm² tissue cultureflask containing 50 ml of growth media and then incubated at 37° C. in a5% CO₂ incubator. The growth media is replaced every 2-3 days until theflask becomes 90% confluent, typically in 5-7 days. To passage andexpand the cell line, a 90% confluent flask is washed with 10 ml of roomtemperature phosphate buffered saline (PBS) and the cells aredisassociated by adding 5 ml 1× Trypsin-EDTA (Invitrogen) and incubatingat 37° C. until the cells detach from the surface of the flask. Toinactivate the trypsin, 5 ml of growth media is added and then thecontents of the flask are centrifuged to pellet the cells. Thesupernatant is aspirated and the cell pellet is resuspended in 10 ml ofgrowth media and the cell number determined using a hemocytometer.Approximately 1-3×10(6) cells per flask are seeded into 175 cm² flaskscontaining 50 ml of growth media and incubated at 37° C. in a 5% CO,incubator. When the flasks reach 90% confluence, the above passagingprocess is repeated until sufficient cells have been obtained forimplantation into mice.

Six to eight week old, female Crl:CD-1-nuBR (nude) mice are obtainedfrom Charles River Laboratories (Wilmington, Mass., USA). Animals arehoused 4-5/cage in micro-isolators, with a 12 hr/12 hr light/dark cycle,acclimated for at least 1 week prior to use and fed normal laboratorychow ad libitum. Studies are conducted on animals between 7 and 12 weeksof age at implantation. To implant tumor cells into nude mice, the cellsare trypsinized as above, washed in PBS and resuspended at aconcentration of 50×10(6) cells/ml in PBS. Using a 27 gauge needle and 1cc syringe, 0.1 ml of the cell suspension is injected into the corpusadiposum of nude mice. The corpus adiposum is a fat body located in theventral abdominal vicera in the right quadrant of the abdomen at thejuncture of the os coxae (pelvic bone) and the os femoris (femur).Tumors are then permitted to develop in vivo until they reachapproximately 150 mm³ in volume, which typically requires 2-3 weeksfollowing implantation. Tumor volumes (V) are calculated by calipermeasurement of the width (W), length (L) and thickness (T) of tumorsusing the following formula: V=0.5326×(L×W×T). Animals are randomizedinto treatment groups so that the average tumor volumes of each groupare similar at the start of dosing.

Sock solutions of test compounds are prepared by dissolving theappropriate amounts of each compound in dimethyl sulfoxide (DMSO) bysonication in an ultrasonic water bath. Stock solutions are prepared atthe start of the study, stored at −20° C. and diluted fresh each day fordosing. A solution of 20% Cremophore RH40 (polyoxyl 40 hydrogenatedcastor oil; BASF Corp., Aktiengesellschaft, Ludwigshafen, Germany) in80% D5W (5% dextrose in water; Abbott Laboratories, North Chicago, Ill.,USA) is also prepared by first heating 100% Cremophore RH40 at 50-60° C.until liquefied and clear, diluting 1:5 with 100% D5W, reheating againuntil clear and then mixing well. This solution is stored at roomtemperature for up to 3 months prior to use. To prepare formulations fordaily dosing, DMSO stock solutions are diluted 1:10 with 20% CremophoreRH40. The final formulation for dosing contains 10% DMSO, 18% CremophoreRH40, 3.6% dextrose and 68.4% water and the appropriate amount of testarticle. Animals are intraperitoneal (IP) injected with this solution at10 ml per kg body weight on a schedule of 5 days per week (Monday thruFriday, with no dosing on Saturday and Sunday) for 3 weeks.

Compounds of the invention are expected to result in decreased thegrowth rate of MDA-MB-435S cells in nude mice to a greater extent than adose of 100 mg/kg body weight of the Hsp90 inhibitor 17-AAG.

Example D Anti-tumor Activity Against Human

Tumor Cells in a nude Mouse Xenograft Model

The human squamous non-small cell lung cancer cell line, RERF-LC-AI(RCB0444; S. Kyoizumi, et al., Cancer. Res. 45:3274-3281, 1985), isobtained from the Riken Cell Bank (Tsukuba, Ibaraki, Japan). The cellline is cultured in growth media prepared from 50% Dulbecco's ModifiedEagle Medium (high glucose), 50% RPMI Media 1640, 10% fetal bovine serum(FBS), 1% 100×L-glutamine, 1% 100× penicillin-streptomycin, 1% 100×sodium pyruvate and 1% 100×MEM non-essential amino acids. FBS isobtained from American Type Culture Collection (Manassas, Va., USA) andall other reagents are obtained from Invitrogen Corp. (Carlsbad, Calif.,USA). Approximately 4-5×10(6) cells that have been cryopreserved inliquid nitrogen are rapidly thawed at 37° C. and transferred to a 175cm² tissue culture flask containing 50 ml of growth media and thenincubated at 37° C. in a 5% CO₂ incubator.

The growth media is replaced every 2-3 days until the flask becomes 90%confluent, typically in 5-7 days. To passage and expand the cell line, a90% confluent flask washed with 10 ml of room temperature phosphatebuffered saline (PBS) and the cells are disassociated by adding 5 ml 1×trypsin-EDTA (Invitrogen) and incubating at 37° C. until the cellsdetach from the surface of the flask. To inactivate the trypsin, 5 ml ofgrowth media is added and then the contents of the flask are centrifugedto pellet the cells. The supernatant is aspirated and the cell pellet isresuspended in 10 ml of growth media and the cell number determinedusing a hemocytometer. Approximately 1-3×10(6) cells per flask areseeded into 175 cm² flasks containing 50 ml of growth media andincubated at 37° C. in a 5% CO₂ incubator. When the flasks reach 90%confluence, the above passaging process is repeated until sufficientcells have been obtained for implantation into mice.

Seven to eight week old, female Crl:CD-1-nuBR (nude) mice are obtainedfrom Charles River Laboratories (Wilmington, Mass., USA). Animals arehoused 4-5/cage in micro-isolators, with a 12 hr/12 hr light/dark cycle,acclimated for at least 1 week prior to use and fed normal laboratorychow ad libitum. Studies are conducted on animals between 8 and 12 weeksof age at implantation. To implant RERF-LC-AI tumor cells into nudemice, the cells are trypsinized as above, washed in PBS and resuspendedat a concentration of 50×10(6) cells/ml in 50% non-supplemented RPMIMedia 1640 and 50% Matrigel Basement Membrane Matrix (#354234; BDBiosciences; Bedford, Mass., USA). Using a 27 gauge needle and 1 ccsyringe, 0.1 ml of the cell suspension is injected subcutaneously intothe flank of each nude mouse. Tumor volumes (V) are calculated bycaliper measurement of the width (W), length (L) and thickness (T) oftumors using the following formula: V=0.5236×(L×W×T).

In vivo passaged RERF-LC-AI tumor cells (RERF-LC-AI^(IVP)) are isolatedto improve the rate of tumor implantation relative to the parental cellline in nude mice. RERF-LC-AI tumors are permitted to develop in vivountil they reach approximately 250 mm³ in volume, which requiresapproximately 3 weeks following implantation. Mice are euthanized viaCO₂ asphyxiation and their exteriors sterilized with 70% ethanol in alaminar flow hood. Using sterile technique, tumors are excised and dicedin 50 ml PBS using a scalpel blade. A single cell suspension is preparedusing a 55 ml Wheaton Safe-Grind tissue grinder (catalog #62400-358; VWRInternational, West Chester, Pa., USA) by plunging the pestle up anddown 4-5 times without twisting. The suspension is strained through a 70μM nylon cell strainer and then centrifuged to pellet the cells. Theresulting pellet is resuspended in 0.1 M NH₄Cl to lyse contaminating redblood cells and then immediately centrifuged to pellet the cells. Thecell pellet is resuspended in growth media and seeded into 175 cm²flasks containing 50 ml of growth media at 1-3 tumors/flask orapproximately 10×10(6) cells/flask. After overnight incubation at 37° C.in a 5% CO₂ incubator, non-adherent cells are removed by rinsing twotimes with PBS and then the cultures are fed with fresh growth media.When the flasks reach 90% confluence, the above passaging process isrepeated until sufficient cells have been obtained for implantation intomice.

RERF-LC-AI^(IVP) cells are then implanted as above and tumors arepermitted to develop in vivo until the majority reached an average of100-200 mm³ in tumor volume, which typically requires 2-3 weeksfollowing implantation. Animals with oblong or very small or largetumors are discarded, and only animals carrying tumors that displayconsistent growth rates are selected for studies. Animals are randomizedinto treatment groups so that the average tumor volumes of each groupare similar at the start of dosing.

The HSP90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG),can be employed as a positive control (Albany Molecular Research,Albany, N.Y., USA). Stock solutions of test articles are prepared bydissolving the appropriate amounts of each compound in dimethylsulfoxide (DMSO) by sonication in an ultrasonic water bath. Stocksolutions are prepared weekly, stored at −20° C. and diluted fresh eachday for dosing. A solution of 20% Cremophore RH40 (polyoxyl 40hydrogenated castor oil; BASF Corp., Aktiengesellschaft, Ludwigshafen,Germany) in 80% D5W (5% dextrose in water; Abbott Laboratories, NorthChicago, Ill., USA) is also prepared by first heating 100% CremophoreRH40 at 50-60° C. until liquefied and clear, diluting 1:5 with 100% D5W,reheating again until clear and then mixing well. This solution isstored at room temperature for up to 3 months prior to use. To prepareformulations for daily dosing, DMSO stock solutions are diluted 1:10with 20% Cremophore RH40. The final formulation for dosing contains 10%DMSO, 18% Cremophore RH40, 3.6% dextrose, 68.4% water and theappropriate amount of test article. Animals are intraperitoneally (i.p.)injected with this solution at 10 ml per kg body weight on a schedule of5 days per week (Monday, Tuesday, Wednesday, Thursday and Friday, withno dosing on Saturday and Sunday) for a total of 15 doses.

Treatment with compounds of the invention is expected to result in thedecreased growth rate of RERF-LC-AI^(IVP) human lung tumor cells in nudemice.

Example E Necrosis in a nude Mouse Tumor Model

The mouse mammary carcinoma cell line, EMT6 (ATCC #CRL-2755), isobtained from the American Type Culture Collection (ATCC; Manassas, Va.,USA). The cell line is cultured in growth media prepared from 50%Dulbecco's Modified Eagle Medium (high glucose), 50% RPMI Media 1640,10% fetal bovine serum (FBS), 1% 100×L-glutamine, 1% 100×Penicillin-Streptomycin, 1% 100× sodium pyruvate and 1% 100×MEMnon-essential amino acids. FBS is obtained from ATCC and all otherreagents are obtained from InVitrogen Corp. (Carlsbad, Calif., USA).

Approximately 4-5×10(6) cells that have been cryopreserved in liquidnitrogen are rapidly thawed at 37° C. and transferred to a 175 cm²tissue culture flask containing 50 ml of growth media and then incubatedat 37° C. in a 5% CO₂ incubator. The growth media is replaced every 2-3days until the flask became 90% confluent, typically in 5-7 days. Topassage and expand the cell line, a 90% confluent flask is washed with10 ml of room temperature phosphate buffered saline (PBS) and the cellsare disassociated by adding 5 ml 1× Trypsin-EDTA (Invitrogen) andincubating at 37° C. until the cells detach from the surface of theflask. To inactivate the trypsin, 5 ml of growth media is added and thenthe contents of the flask are centrifuged to pellet the cells. Thesupernatant is aspirated and the cell pellet is resuspended in 10 ml ofgrowth media and the cell number determined using a hemocytometer.Approximately 1-3×10(6) cells per flask are seeded into 175 cm² flaskscontaining 50 ml of growth media and incubated at 37° C. in a 5% CO₂incubator. When the flasks reach 90% confluence, the above passagingprocess is repeated until sufficient cells have been obtained forimplantation into mice.

Seven to eight week old, female Crl:CD-1-nuBR (nude) mice are obtainedfrom Charles River Laboratories (Wilmington, Mass., USA). Animals arehoused 4-5/cage in micro-isolators, with a 12 hr/12 hr light/dark cycle,acclimated for at least 1 week prior to use and fed normal laboratorychow ad libitum. Studies are conducted on animals between 8 and 10 weeksof age at implantation. To implant EMT6 tumor cells into nude mice, thecells are trypsinized as above, washed in PBS and resuspended at aconcentration of 10×10(6) cells/ml in PBS. Using a 27 gauge needle and 1cc syringe, 0.1 ml of the cell suspension is injected subcutaneouslyinto the flank of each nude mouse.

Tumors are then permitted to develop in vivo until the majority reached75-125 mm³ in tumor volume, which typically requires 1 week followingimplantation. Animals with oblong, very small or large tumors arediscarded, and only animals'carrying tumors that display consistentgrowth rates are selected for studies. Tumor volumes (V) are calculatedby caliper measurement of the width (W), length (L) and thickness (T) oftumors using the following formula: V=0.5236×(L×W×T). Animals arerandomized into treatment groups so that each group had median tumorvolumes of ˜100 mm³ at the start of dosing.

To formulate a compound of the invention in DRD, a stock solution of thetest article is prepared by dissolving an appropriate amount of thecompound in dimethyl sulfoxide (DMSO) by sonication in an ultrasonicwater bath, A solution of 20% Cremophore RH40 (polyoxyl 40 hydrogenatedcastor oil; BASF Corp., Aktiengesellschaft, Ludwigshafen, Germany) in 5%dextrose in water (Abbott Laboratories, North Chicago, Ill., USA) isalso prepared by first heating 100% Cremophore RH40 at 50-60° C. untilliquefied and clear, diluting 1:5 with 100% D5W, reheating again untilclear and then mixing well. This solution is stored at room temperaturefor up to 3 months prior to use. To prepare a DRD formulation fordosing, the DMSO stock solution is diluted 1:10 with 20% CremophoreRH40. The final DRD formulation for dosing contains 10% DMSO, 18%Cremophore RH40, 3.6% dextrose, 68.4% water and the appropriate amountof test article.

Tumor-bearing animals are given a single intravenous (i.v.) bolusinjections of either DRD vehicle or a compound of the inventionformulated in DRD, both at 10 mL per kg body weight. Then, 4-24 hr afterdrug treatment, tumors are excised, cut in half and fixed overnight in10% neutral-buffered formalin. Each tumor is embedded in paraffin withthe cut surfaces placed downwards in the block, and rough cut until acomplete section is obtained. From each tumor, 5 μM serial sections areprepared and stained with hematoxylin and eosin. Slides are evaluatedmanually using light microscopy with a 10×10 square gridded reticle. Thepercentage of necrosis in a tumor is quantified at 200× magnification byscoring the total number of grid squares containing necrosis and thetotal number of grid squares containing viable tumor cells.

It is expected that compounds of the invention will result in anincrease in necrotic tissue in the center of EMT6 tumors relative to thebaseline necrosis observed in vehicle treated tumors. As would beexpected for a vascular targeting mechanism of action, rapid onset ofnecrosis is consistent with there being a loss of blood flow to tumorsresulting in hypoxia and tumor cell death.

Example F Vascular Disrupting Activities in a nude Mouse Tumor Model

The mouse mammary carcinoma cell line, EMT6 (ATCC #CRL-2755), isobtained from the American Type Culture Collection (ATCC; Manassas, Va.,USA). The cell line is cultured in growth media prepared from 50%Dulbecco's Modified Eagle Medium (high glucose), 50% RPMI Media 1640,10% fetal bovine serum (FBS), 1% 100×L-glutamine, 1% 100×Penicillin-Streptomycin, 1% 100)C sodium pyruvate and 1% 100×MEMnon-essential amino acids. FBS is obtained from ATCC and all otherreagents are obtained from Invitrogen Corp. (Carlsbad, Calif., USA).Approximately 4-5×10⁶ cells that have been cryopreserved in liquidnitrogen are rapidly thawed at 37° C. and transferred to a 175 cm²tissue culture flask containing 50 mL of growth media and then incubatedat 37° C. in a 5% CO₂ incubator. The growth media is replaced every 2-3days until the flask became 90% confluent, typically in 5-7 days. Topassage and expand the cell line, a 90% confluent flask is washed with10 mL of room temperature phosphate buffered saline (PBS) and the cellsare disassociated by adding 5 mL 1× Trypsin-EDTA (Invitrogen) andincubating at 37° C. until the cells detach from the surface of theflask. To inactivate the trypsin, 5 mL of growth media is added and thenthe contents of the flask are centrifuged to pellet the cells. Thesupernatant is aspirated and the cell pellet is resuspended in 10 mL ofgrowth media and the cell number determined using a hemocytometer.Approximately 1-3×10⁶ cells per flask are seeded into 175 cm² flaskscontaining 50 mL of growth media and incubated at 37° C. in a 5% CO₂incubator. When the flasks reach 90% confluence, the above passagingprocess is repeated until sufficient cells have been obtained forimplantation into mice.

Seven to eight week old, female Crl:CD-1-nuBR (nude) mice are obtainedfrom Charles River Laboratories (Wilmington, Mass., USA). Animals arehoused 4-5/cage in micro-isolators, with a 12 hr/12 hr light/dark cycle,acclimated for at least 1 week prior to use and fed normal laboratorychow ad libitum. Studies are conducted on animals between 8 and 10 weeksof age at implantation. To implant EMT6 tumor cells into nude mice, thecells are trypsinized as above, washed in PBS and resuspended at aconcentration of 10×10⁶ cells/mL in PBS. Using a 27 gauge needle and 1cc syringe, 0.1 mL of the cell suspension is injected subcutaneouslyinto the flank of each nude mouse.

For the Evans Blue dye assay, tumors are permitted to develop in vivountil the majority reach 40-90 mm³ in tumor volume (to minimize theextent of tumor necrosis), which typically require 4-6 days followingimplantation. Animals with visibly necrotic, oblong, very small or verylarge tumors are discarded and only animals carrying tumors that displayconsistent growth rates are selected for use. Tumor volumes (V) arecalculated by caliper measurement of the width (W), length (L) andthickness (T) of tumors using the following formula: V=0.5236×(L×W×T).Animals are randomized into treatment groups so that at the start ofdosing each group have median tumor volumes of ˜125 mm³ or ˜55 mm³ forthe Evans Blue dye assay.

To formulate compounds of the invention for dosing, the appropriateamount of compound is dissolved in 5% dextrose in water (D5W; AbbottLaboratories, North Chicago, Ill., USA). Vehicle-treated animals aredosed with D5W.

To conduct the Evans Blue dye assay, tumor-bearing animals are dosedwith vehicle or test article at 0 hr, and then i.v. injected with 100 μLof a 1% (w/v) Evan's Blue dye (Sigma #E-2129; St. Louis, Mo., USA)solution in 0.9% NaCl at +1 hr. Tumors are excised at +4 hr, weighed andthe tissue disassociated by incubation in 50 μL 1 N KOH at 60° C. for 16hr. To extract the dye, 125 μL of a 0.6 N phosphoric acid and 325 μLacetone are added, and the samples vigorously vortexed and thenmicrocentrifuged at 3000 RPM for 15 min to pellet cell debris. Theoptical absorbance of 200 μL of supernatant is then measured at 620 nMin a Triad spectrophotometer (Dynex Technologies, Chantilly, Va., USA).Background OD₆₂₀ values from similarly sized groups of vehicle or testarticle-treated animals that have not been injected with dye aresubtracted as background. OD₆₂₀ values are then normalized for tumorweight and dye uptake is calculated relative to vehicle-treated tumors.

To examine the vascular disrupting activity of a compound of theinvention, the Evans Blue dye assay is employed as a measurement oftumor blood volume (Graff et al., Eur J Cancer 36:1433-1440, 2000).Evans Blue dye makes a complex with serum albumin by electrostaticinteraction between the sulphonic acid group of the dye and the terminalcationic nitrogens of the lysine residues in albumin. The dye leaves thecirculation very slowly, principally by diffusion into extravasculartissues while still bound to albumin. Albumin-dye complex taken up bytumors is located in the extracellular space of non-necrotic tissue, andintracellular uptake and uptake in necrotic regions is negligible. Theamount of dye present in a tumor is a measurement of the tumor bloodvolume and microvessel permeability. Compounds of the invention areexpected to result in substantially decreased tumor dye uptake relativeto vehicle-treated animals. Such a decrease in dye penetration into thetumor is consistent with there being a loss of blood flow to tumors dueto blockage of tumor vasculature, consistent with a vascular disruptingmechanism of action.

Example G Inhibition of the Production of Inflammatory Cytokines inHuman PBMCs

Human PBMC are isolated using Ficoll 400 and diatrizoate sodium (density1.077 g/ml) solution and purified with RosetteSep (StemCellTechnologies). The PBMCs are primed with human IFN-γ (800 U/ml, PierceBiotechnology #R—IFNG-50), seeded at 0.5×10⁶/100μL/well in 96-wellU-bottom plate with culture medium (RPMI 1640, 10% FBS, 1% Pen/Strep),and incubated in 37° C. for overnight. The cells are then stimulatedwith 1 μg/ml of LPS (Lipopolysaccharide, Sigma#L2654-1MG) or 0.025% ofSAC (Staphylococcus Aureus Cowan, Calbiochem-Novabiochem Corp. #507858),and treated with a test compound at different concentrations with finalDMSO concentration less than 0.5% for 16-18 hrs. About 180411/well ofsupernatant is 15, collected and measured using ELISA kit or Bio-plex(Bio-Rad) to determine the levels of cytokine production. The cellsurvival is determined using Cell Counting Kit-8 (Dojindo MolecularTechnologies, Inc.). Compounds of the invention are expected to broadlyinhibit the production of proinflammatory cytokines.

Example H Suppression of Glucocorticoid Receptor Levels in Rat and HumanPBMCs Cell Preparation:

Whole blood samples from healthy human volunteers and male SD rats arecollected and the PBMCs are isolated immediately as follows. 5 ml ofwhole blood is diluted with an equal volume of sterile 1×PBS. Thediluted blood is overlayed carefully into a sterile centrifuge tubewithout disturbing the bottom layer that containing 5 ml of Ficoll-paqueplus density gradient solution. The layered blood is centrifuged at1500×g for 30 minutes at room temperature. The middle thin layercontaining PBMCs is carefully removed, transferred to another sterilecentrifuge tube, and washed twice with PBS to remove Percoll. Isolatedrat and human PBMCs are cultured in 10% fetal bovine serum/DMEM.

Treatment:

The rat and human PBMCs are treated with DMSO (control), compounds ofthe invention, or 17-DMAG at concentrations of 0, 1, 5, 25, or 100 nM(in DMSO) for 16 hours. The cells are then collected and rinsed inice-cold PBS and stored in liquid nitrogen until further analysis.

Immunoblot

PBMC are prepared in Western lysis buffer (10 mmol/LHEPES, 42 mmol/LKCl, 5 mmol/L MgCl₂, 0.1 mmol/L EDTA, 0.1 mmol/L EGTA, 1 mmol/L DTT, 1%Triton X-100, freshly supplemented with 1× protease inhibitor cocktailfrom Pierce, Rockford, Ill.). Lysate protein concentrations arequantified by bicinchoninic acid assay (Pierce) and normalized. Equalamounts of protein are loaded onto 10% NuPAGE Bis-Tris Gels (Invitrogen)and subsequently transferred onto polyvinylidene difluoride membranes.The membranes are blocked in 5% milk in TBST. Primary antibody ofglucocorticod receptor from Santa Cruz Biotechnology, Inc. is added andincubated at room temperature for 1 hour with shaking. The blots arewashed extensively in TBST before secondary antibodies are added forovernight incubation at 4° C. with gentle shaking. The blots are againwashed extensively and developed with SuperSignal West Femto substrate(Pierce). The immunoblot analysis is performed to measure the level oftotal GRs by Quantity One software from Bio-Rad.

Example I Suppression of Glucocorticoid Receptor Levels in Human PBMCsand Renal Cells, as well as in Several Human Cancer Cell Lines CellPreparation:

Normal human renal proximal tubule epithelial cells and tumor cell linesof MV-4-11, Kasumi-1, and Hela are obtained from Cambrex Bioproducts andAmerican Type Culture Collection, respectively. Cells are cultured with10% fetal bovine serum/DMEM.

The whole blood samples from healthy human volunteers are collected andthe PBMCs are isolated immediately as described in Example H. Isolatedhuman PBMCs are cultured in 10% fetal bovine serum/DMEM.

Treatment:

Human PBMCs, kasumi-1, Mv-4-11, Hela, and human renal proximal tubuleepithelial cells are treated with DMSO (control), compounds of theinvention, 17-DMAG at concentrations of 0, 5, 25, or 100 nM (in DMSO)for 16 hours. The cells are then collected and rinsed in ice-cold PBSand stored in liquid nitrogen until further analysis.

Immunoblot

PBMC, renal and tumor cell pellets are prepared in Western lysis buffer(10 mmol/L HEPES, 42 mmol/L KCl, 5 mmol/L MgCl₂, 0.1 mmol/L EDTA, 0.1mmol/L EGTA, 1 mmol/L DTT, 1% Triton X-100, freshly supplemented with 1×protease inhibitor cocktail from Pierce, Rockford, Ill.). Lysate proteinconcentrations are quantified by bicinchoninic acid assay (Pierce) andnormalized. Equal amounts of protein are loaded onto 10% NuPAGE Bis-TrisGels (Invitrogen) and subsequently transferred onto polyvinylidenedifluoride membranes. The membranes are blocked in 5% milk in TBST.Primary antibody of glucocorticod receptor from Santa CruzBiotechnology, Inc. is added and incubated at room temperature for 1hour with shaking. The blots are washed extensively in TBST beforesecondary antibodies are added for overnight incubation at 4° C. withgentle shaking. The blots are again washed extensively and developedwith SuperSignal West Femto substrate (Pierce). Compounds of theinvention are expected to suppress the expression of glucocorticoidreceptors in cancer cells as well as in normal PBMCs and renal cells.

Example J Suppression of Glucocorticoid Receptor Levels In vivo

Male adult Sprague-Dawley (SD) rats, five per group, are randomlyassigned into five testing groups which receive treatments as shown inTable 5:

TABLE 5 Treatment group Treatment G1 5 mL/kg of vehicle (5% DMSO/13.5%Cr-RH40/D5W) G2 6 mg/kg of 17-DMAG G3 5 mg/kg of Paclitaxel G4 80 mg/kgof Compound of the invention G5 50 mg/kg of Compound of the invention

The test compounds are administered daily intravenously via tail veinfor four days. All rats are sacrificed at the study day 5. About 1-2 mLof blood samples are collected per animal. The blood samples are thenpulled together as a group for PBMC isolation. PBMCs are isolated and animmunoblot using an antibody that recognizes the glucocorticoid receptoris prepared, as described in Examples H and I.

Example K Inhibition of Topoisomerase II

The ability of compounds of the invention to inhibit the activity oftopoisomerase II is examined with a kDNA decatenation assay (TopoGEN,Inc. Port Orange, Fla.). Substrate kDNA is mixed with compounds (10,100, or 500 μM) and incubated at 37° C. for 30 min. The reaction is stopby adding ⅕ volume of stop buffer. 20 μl of the reaction is loaded on 1%agarose gel. Image of decatenation of kDNA by compounds is taken byKodak Image Station 440.

All publications, patent applications, patents, and other documentscited herein are incorporated by reference in their entirety. In case ofconflict, the present specification, including definitions, willcontrol. In addition, the materials, methods, and examples areillustrative only and not intended to be limiting.

Example L Compound #3 Displays Anti-tumor Activity Against Human andCanine Tumor Cells in SCID Mouse Xenograft Models

The human multiple myeloma cell line, RPMI 8226 (ATCC #CCL-155), and thecanine osteosarcoma cell line, D-17 (ATCC #CCL-183), were obtained fromthe American Type Culture Collection (ATCC; Manassas, Va., USA). Thecells were cultured in growth media prepared with RPMI Media 1640 (4.5g/L glucose), 10% fetal bovine serum (FBS), 10 in M HEPES, 1% 100×Penicillin-Streptomycin, 1% 100× sodium pyruvate and 1% 100×MEMnon-essential amino acids. FBS was obtained from ATCC and all otherreagents were obtained from Invitrogen Corp. (Carlsbad, Calif., USA).Cells that had been cryopreserved in liquid nitrogen were rapidly thawedat 37° C. and transferred to a tissue culture flask containing growthmedia and then incubated at 37° C. in a 5% CO₂ incubator. To expand theRPMI 8226 cell line, growth media was changed every 2-3 days andcultures were passaged 1:3 to 1:5 every 3-5 days. When the a 175 cm²flask reached approximately 20-40×10(6) total cells, the above passagingprocess was repeated until sufficient cells had been obtained forimplantation into mice. To expand the D-17 cell line, cultures weresplit 1:4 every 2 days when 175 cm² flasks became 75% confluent. D-17cultures were passaged by washing with 10 mL of room temperaturephosphate buffered saline (PBS) and then disassociating cells by adding5 mL 1× trypsin-EDTA and incubating at 37° C. until the cells detachedfrom the surface of the flask. To inactivate the trypsin, 5 mL of growthmedia was added and then the contents of the flask were centrifuged topellet the cells. The supernatant was aspirated and the cell pellet wasresuspended in 10 mL of growth media and the cell number determinedusing a hemocytometer. Cells were seeded into 175 cm² flasks containing50 mL of growth media and incubated at 37° C. in a 5% CO₂ incubator.When the flasks reached 75% confluence, the above passaging process wasrepeated until sufficient cells had been obtained for implantation intomice.

Seven to eight week old, female CB17/Icr-Prkdc^(scid)/Crl (SCID) micewere obtained from Charles River Laboratories (Wilmington, Mass., USA).Animals were housed 4-5/cage in micro-isolators, with a 12 hr/12 hrlight/dark cycle, acclimated for at least 1 week prior to use and fednormal laboratory chow ad libitum. Animals were between eight to nineweeks of age at implantation. To implant RPMI 8226 and D-17 tumor cellsinto SCID mice, cells were collected as described above, washed in PBSand resuspended at a concentration of 5×10(7) cells/mL in 50%non-supplemented medium and 50% Matrigel Basement Membrane Matrix(#354234; BD Biosciences; Bedford, Mass., USA). Using a 27 gauge needleand 1 cc syringe, 5×10(6) RPMI 8226 or D-17 cells in 0.1 mL of a cellsuspension were injected subcutaneously into the flanks of SCID mice.

Tumors were then permitted to develop in vivo until the majority reached75-250 mm³ in tumor volume, which required ˜4 weeks followingimplantation for the RPMI 8226 model and ˜9 weeks for the D-17 model.Animals with oblong, very small or large tumors were discarded and onlyanimals carrying tumors that displayed consistent growth rates wereselected for studies. Tumor volumes (V) were calculated by calipermeasurement of the width (W), length (L) and thickness (T) of tumorsusing the following formula: V=0.5236×(L×W×T). Animals were randomizedinto treatment groups so that the average tumor volumes of each groupwere similar at the start of dosing. % T/C values, as a measure ofefficacy, were determined as follows:

-   -   (i) If ΔT>0: % T/C=(ΔT/ΔC)×100    -   (ii) If ΔT<0: % T/C=(ΔT/T₀)×100    -   (iii) ΔT=Change in average tumor volume between start of dosing        and the end of study.    -   (iv) ΔC=Change in average tumor volume between start of dosing        and the end of study.    -   (v) T₀=Average tumor volume at start of dosing.

To formulate Compound #3 in DRD, stock solutions of the test articlewere prepared by dissolving the appropriate amounts of the compound indimethyl sulfoxide (DMSO) by sonication in an ultrasonic water bath.Stock solutions were prepared weekly, stored at −20° C. and dilutedfresh each day for dosing. A solution of 20% Cremophore RH40 (polyoxyl40 hydrogenated castor oil; BASF Corp., Aktiengesellschaft,Ludwigshafen, Germany) in 5% dextrose in water (Abbott Laboratories,North Chicago, Ill., USA) was also prepared by first heating 100%Cremophore RH40 at 50-60° C. until liquefied and clear, diluting 1:5with 100% D5W, reheating again until clear and then mixing well. Thissolution can be stored at room temperature for up to 3 months prior touse. To prepare DRD formulations for daily dosing, DMSO stock solutionswere diluted 1:10 with 20% Cremophore RH40. The final DRD formulationfor dosing contained 10% DMSO, 18% Cremophore RH40, 3.6% dextrose, 68.4%water and the appropriate amount of test article. Animals wereintravenously (i.v.) injected with this formulation at 10 mL per kg bodyweight on either 1 day each week or 3 days each week (Monday, Wednesday,Friday).

FIG. 1 is a plot showing an average tumor volume, as a function of dayspost-implantation, of a SCID mouse xenograft study that determined theeffects of Compound #3 on the in vivo growth rate of the human multiplemyeloma tumor cell line RPMI 8226. As can be seen from the plot,treatment with a dose of 100 mg/kg body weight of Compound #3substantially inhibited tumor growth, with a % T/C value of 15 observedon day 46. Overt toxicity was not observed, with the 100 mg/kg Compound#3-treated group having an average bodyweight change on day 46 relativeto the start of the study of 2.1% (+/−0.9 SEM), as compared to 2.6%(+/−3.6 SEM) for the vehicle-treated group.

FIG. 2 is a plot showing an average tumor volume, as a function of dayspost-implantation, of a SCID mouse xenograft study to determine theeffects of Compound #3 on the in vivo growth rate of the canineosteosarcoma tumor cell line D17. As can be seen from the plt, treatmentwith a dose of 61 mg/kg body weight of Compound #3 substantiallyinhibited tumor growth, with a % T/C value of 6 observed on day 83.Overt toxicity was not observed, with the 61 mg/kg Compound #3-treatedgroup having an average bodyweight change on day 83 relative to thestart of the study of −0.2% (+/−1.9 SEM), as compared to 0.4% (+/−1.0SEM) for the vehicle-treated group.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by the following structural formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or clathratethereof, wherein: R₁, R₂ and R₃ are independently —OH, —SH, —NR₇H,—OR₂₆, —SR₂₆, —O(CH₂)_(m)OH, —O(CH₂)_(m)SH, —O(CH₂)_(m)NR₇H,—S(CH₂)_(m)OH, —S(CH₂)_(m)SH, —S(CH₂)_(m)NR₇H, —OC(O)NR₁₀R₁₁,—SC(O)NR₁₀R₁₁, —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇,—OC(O)OR₇, —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇,—NR₇CH₂C(O)R₇, —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇,—OCH₂C(O)NR₁₀R₁₁, —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,—SS(O)_(p)R₇, —S(O)_(p)OR₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁,—SS(O)_(p)NR₁₀R₁₁, —NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇, —SS(O)_(p)OR₇,—NR₇S(O)_(p)OR₇, —OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇, —OC(S)OR₇, —SC(S)OR₇,—NR₇C(S)OR₇, —OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁, —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇,—SC(NR₈)R₇, —NR₇C(NR₈)R₇, —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇,—OC(NR₈)NR₁₀R₁₁, —SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —OP(O)(OR₇)₂, or—SP(O)(OR₇)₂, provided that at least one of R₁, R₂ and R₃ is—OP(O)(OH)₂; R₅ is —X₂₀R₅₀, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl; R₇ andR₈, for each occurrence, is independently, —H, an optionally substitutedalkyl, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl; R₁₀ andR₁₁, for each occurrence, is independently —H, an optionally substitutedalkyl, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl; or R₁₀and R₁₁, taken together with the nitrogen to which they are attached,form an optionally substituted heterocyclyl or an optionally substitutedheteroaryl; R₂₆ is a lower alkyl; R₅₀ is an optionally substituted arylor an optionally substituted heteroaryl; X₂₀ is a C1-C4 alkyl, NR₇,C(O), C(S), C(NR₈), or S(O)_(p); p, for each occurrence, isindependently, 1 or 2; m for each occurrence, is independently 1, 2, 3,or 4; n is 0, 1, 2, or 3; and Z, for each occurrence, is independentlyan optionally substituted alkyl, an optionally substituted alkenyl, anoptionally substituted alkynyl, an optionally substituted cycloalkyl, anoptionally substituted cycloalkenyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, an optionally substitutedheteroaralkyl, halo, cyano, nitro, guanadino, a haloalkyl, aheteroalkyl, alkoxy, haloalkoxy, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇,—C(S)R₇, —C(O)SR₇, —C(S)SR₇, —C(S)OR₇, —C(S)NR₁₀R₁₁, —C(NR₈)OR₇,—C(NR₈)R₇, —C(NR₈)NR₁₀R₁₁, —C(NR₈)SR₇, —OC(O)R₇, —OC(O)OR₇, —OC(S)OR₇,—OC(NR₈)OR₇, —SC(O)R₇, —SC(O)OR₇, —SC(NR₈)OR₇, —OC(S)R₇, —SC(S)R₇,—SC(S)OR₇, —OC(O)NR₁₀OR₁₁, —OC(S)NR₁₀OR₁₁, —OC(NR₈)NR₁₀OR₁₁,—SC(O)NR₁₀OR₁₁, —SC(NR₈)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇,—C(O)NR₁₀R₁₁, —NR₈C(O)R₇, —NR₇C(S)R₇, —NR₇C(S)OR₇, —NR₇C(NR₈)R₇,—NR₇C(O)OR₇, —NR₇C(NR₈)OR₇, —NR₇C(O)NR₁₀OR₁₁, —NR₇C(S)NR₁₀OR₁₁,—NR₇C(NR₈)NR₁₀OR₁₁, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇, —OS(O)_(p)OR₇,—OS(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —NR₈S(O)_(p)R₇, —NR₇S(O)_(p)NR₁₀OR₁₁,—NR₇S(O)_(p)OR₇, —S(O)_(p)NR₁₀R₁₁, —SS(O)_(p)R₇, —SS(O)OR₇,—SS(O)_(p)NR₁₀R₁₁, —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂; and with the provisothat the compound is not3-hydroxy-4-(5-mercapto-4-(naphthalen-1-yl)-4H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate. 2-13. (canceled)
 14. The compound of claim 1,wherein Z is a C1-C6 alkyl, a C1-C6 haloalkyl, a C1-C6 alkoxy, a C1-C6haloalkoxy, a C1-C6 alkyl sulfanyl or a C3-C6 cycloalkyl; and n is 1.15-19. (canceled)
 20. The compound of claim 1, represented by thefollowing structural formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or clathratethereof, wherein: R₆ is an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, cyano, halo,nitro, an optionally substituted cycloalkyl, haloalkyl, an optionallysubstituted heterocyclyl, an optionally substituted aryl, an optionallysubstituted heteroaryl, an optionally substituted aralkyl, an optionallysubstituted heteroaralkyl, —OR₇, —SR₇, —NR₁₀R₁₁, —OC(O)NR₁₀R₁₁,—SC(O)NR₁₀R₁₁, —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇,—OC(O)OR₇, —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇,—NR₇CH₂C(O)R₇, —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇,—OCH₂C(O)NR₁₀R₁₁, —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,—SS(O)_(p)R₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁, —SS(O)_(p)NR₁₀R₁₁,—NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇, —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇,—OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇, —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇,—OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁, —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇,—NR₇C(NR₈)R₇, —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,—SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —C(O)R₇, —C(O)OR₇, —C(O)NR₁₀R₁₁,—C(O)SR₇, —C(S)R₇, —C(S)OR₇, —C(S)NR₁₀R₁₁, —C(S)SR₇, —C(NR₈)OR₇,—C(NR₈)R₇, —C(NR₈)NR₁₀R₁₁, —C(NR₈)SR₇, —S(O)_(p)OR₇, —S(O)_(p)NR₁₀R₁₁,or —S(O)_(p)R₇. 21-33. (canceled)
 34. The compound of claim 20, whereinone of R₁, R₂, or R₃ is —OP(O)(OH)₂. 35-36. (canceled)
 37. The compoundof claim 1, represented by the following structural formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or clathratethereof.
 38. The compound of claim 37, wherein R₅ is represented by thefollowing formula:

wherein: R₂₇, for each occurrence, is independently a substituentselected from the group consisting of —H, an optionally substitutedalkyl, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, an optionally substituted heteraralkyl, halo,cyano, nitro, guanadino, a hydroxyalkyl, alkoxyalkyl, haloalkyl, aheteroalkyl, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁,—NR₈C(O)R₇, —SR₇, —S(O)_(p)OR₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,—NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —OP(O)(OR₇)₂,—SP(O)(OR₇)₂, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂; or two R₂₇groups taken together with the carbon atom to which they are attachedform an optionally substituted cycloalkyl or optionally substitutedheterocyclyl ring; R₉, for each occurrence, is independently asubstituent selected from the group consisting of an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, an optionally substituted heteraralkyl,hydroxyalkyl, alkoxyalkyl, halo, cyano, nitro, guanadino, a haloalkyl, aheteroalkyl, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁,—NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,—NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or—SP(O)(OR₇)₂; or two R₉ groups taken together with the carbon atoms towhich they are attached form a fused ring; and q is zero or an integerfrom 1 to
 7. 39. (canceled)
 40. The compound of claim 37, wherein R₅ isrepresented by the following formula:

wherein: R₃₃ is a halo, lower alkyl, a lower alkoxy, a lower haloalkyl,a lower haloalkoxy, and lower alkyl sulfanyl; R₃₄ is H, a lower alkyl,or a lower alkylcarbonyl; and Ring B and Ring C are optionallysubstituted with one or more substituents.
 41. The compound of claim 37,wherein R₅ is selected from the group consisting of:

wherein: X₆, for each occurrence, is independently CH, CR₉, N,N(O),N⁺(R₁₇), provided that at least three X₆ groups are independentlyselected from CH and CR₉; X₇, for each occurrence, is independently CH,CR₉, N,N(O), N⁺(R₁₇), provided that at least three X₇ groups areindependently selected from CH and CR₉; X₈, for each occurrence, isindependently CH₂, CHR₉, C(R₉)₂, S, S(O)_(p), NR₇, or NR₁₇; X₉, for eachoccurrence, is independently N or CH; X₁₀, for each occurrence, isindependently CH, CR₉, N,N(O), N⁺(R₁₇), provided that at least one X₁₀is selected from CH and CR₉; R₉, for each occurrence, is independently asubstituent selected from the group consisting of an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, an optionally substituted heteraralkyl,hydroxyalkyl, alkoxyalkyl, halo, cyano, nitro, guanadino, a haloalkyl, aheteroalkyl, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁,—NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,—NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or—SP(O)(OR₇)₂, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂; or two R₉groups taken together with the carbon atoms to which they are attachedform a fused ring; and R₁₇, for each occurrence, is independently —H, analkyl, an aralkyl, —C(O)R₇, —C(O)OR₇, or —C(O)NR₁₀R₁₁.
 42. The compoundof claim 41, wherein R₅ is an optionally substituted indolyl, anoptionally substituted benzoimidazolyl, an optionally substitutedindazolyl, an optionally substituted 3H-indazolyl, an optionallysubstituted indolizinyl, an optionally substituted quinolinyl, anoptionally substituted isoquinolinyl, an optionally substitutedbenzoxazolyl, an optionally substituted benzo[1,3]dioxolyl, anoptionally substituted benzofuryl, an optionally substitutedbenzothiazolyl, an optionally substituted benzo[d]isoxazolyl, anoptionally substituted benzo[d]isothiazolyl, an optionally substitutedthiazolo[4,5-c]pyridinyl, an optionally substitutedthiazolo[5,4-c]pyridinyl, an optionally substitutedthiazolo[4,5-b]pyridinyl, an optionally substitutedthiazolo[5,4-b]pyridinyl, an optionally substitutedoxazolo[4,5-c]pyridinyl, an optionally substitutedoxazolo[5,4-c]pyridinyl, an optionally substitutedoxazolo[4,5-b]pyridinyl, an optionally substitutedoxazolo[5,4-b]pyridinyl, an optionally substituted imidazopyridinyl, anoptionally substituted benzothiadiazolyl, benzoxadiazolyl, an optionallysubstituted benzotriazolyl, an optionally substituted tetrahydroindolyl,an optionally substituted azaindolyl, an optionally substitutedquinazolinyl, an optionally substituted purinyl, an optionallysubstituted imidazo[4,5-a]pyridinyl, an optionally substitutedimidazo[1,2-a]pyridinyl, an optionally substituted3H-imidazo[4,5-b]pyridinyl, an optionally substituted1H-imidazo[4,5-b]pyridinyl, an optionally substituted1H-imidazo[4,5-c]pyridinyl, an optionally substituted3H-imidazo[4,5-c]pyridinyl, an optionally substituted pyridopyrdazinyl,and optionally substituted pyridopyrimidinyl, an optionally substitutedpyrrolo[2,3]pyrimidyl, an optionally substituted pyrazolo[3,4]pyrimidylan optionally substituted cyclopentaimidazolyl, an optionallysubstituted cyclopentatriazolyl, an optionally substitutedpyrrolopyrazolyl, an optionally substituted pyrroloimidazolyl, anoptionally substituted pyrrolotriazolyl, or an optionally substitutedbenzo[b]thienyl.
 43. The compound of claim 37, wherein R₅ is selectedfrom the group consisting of:

wherein: X₁₁, for each occurrence, is independently CH, CR₉, N,N(O), orN⁺(R₁₇); X₁₂, for each occurrence, is independently CH, CR₉, N,N(O),N⁺(R₁₇), provided that at least one X₁₂ group is independently selectedfrom CH and CR₉; X₁₃, for each occurrence, is independently O, S,S(O)_(p), NR₇, or NR₁₇; R₉, for each occurrence, is independently asubstituent selected from the group consisting of an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, an optionally substituted heteraralkyl,halo, cyano, nitro, guanadino, a hydroxyalkyl, alkoxyalkyl, haloalkyl, aheteroalkyl, —NR₁₀R₁₁, —OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁,—NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,—NR₈S(O)_(p)R₇, or —S(O)_(p)NR₁₀R₁₁, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or—SP(O)(OR₇)₂, —S(O)_(p)OR₇, —OP(O)(OR₇)₂, or —SP(O)(OR₇)₂; or two R₉groups taken together with the carbon atoms to which they are attachedform a fused ring; and R₁₇, for each occurrence, is independently analkyl or an aralkyl. 44-46. (canceled)
 47. The compound of claim 37,wherein R₅ is X₂₀R₅₀; X₂₀ is a C1-C4 alkyl and R₅₀ is an optionallysubstituted phenyl.
 48. (canceled)
 49. The compound of claim 37, whereinR₆ is a C1-C6 alkyl, a C1-C6 haloalkyl, a C1-C6 alkoxy, a C1-C6haloalkoxy, a C1-C6 alkyl sulfanyl or a C3-C6 cycloalkyl.
 50. Thecompound of claim 49, wherein R₆ is selected from the group consistingof methyl, ethyl, isopropyl, and cyclopropyl.
 51. The compound of claim37, wherein R₁ and R₃ are each independently —OH, —SH, or —NHR₇.
 52. Thecompound of claim 51, wherein R₁ and R₃ are both —OH. 53-66. (canceled)67. The compound of claim 1, represented by the following structuralformula:

or a tautomer, pharmaceutically acceptable salt solvate, or clathratethereof; wherein: X₄₁ is O, S, or NR₄₂; X₄₂ is CR₄₄ or N; Y₄₀ is N orCR₄₃; Y₄₁ is N or CR₄₅; Y₄₂, for each occurrence, is independently N, Cor CR₄₆; R₄₁ is an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, cyano, halo,nitro, an optionally substituted cycloalkyl, haloalkyl, an optionallysubstituted heterocyclyl, an optionally substituted aryl, an optionallysubstituted heteroaryl, an optionally substituted aralkyl, an optionallysubstituted heteroaralkyl, —OR₇, —SR₇, —NR₁₀R₁₁, —OC(O)NR₁₀R₁₁,—SC(O)NR₁₀OR₁₁, —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇, —NR₇C(O)R₇,—OC(O)OR₇, —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇,—NR₇CH₂C(O)R₇, —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇,—OCH₂C(O)NR₁₀R₁₁, —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,—SS(O)_(p)R₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀OR₁₁, —SS(O)_(p)NR₁₀R₁₁,—NR₇S(O)_(p)NR₁₀OR₁₁, —OS(O)_(p)OR₇, —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇,—OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇, —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇,—OC(S)NR₁₀R₁₁, —SC(S)NR₁₀R₁₁, —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇,—NR₇C(NR₈)R₇, —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,—SC(NR₈)NR₁₀R₁₁, —NR₇C(NR₈)NR₁₀R₁₁, —C(O)R₇, —C(O)OR₇, —C(O)NR₁₀R₁₁,—C(O)SR₇, —C(S)R₇, —C(S)OR₇, —C(S)NR₁₀OR₁₁, —C(S)SR₇, —C(NR₈)OR₇,—C(NR₈)R₇, —C(NR₈)NR₁₀R₁₁, —C(NR₈)SR₇, —S(O)_(p)OR₇, —S(O)_(p)NR₁₀OR₁₁,or —S(O)_(p)R₇; R₄₂ is —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, an optionally substituted heteraralkyl,hydroxyalkyl, alkoxyalkyl, a haloalkyl, a heteroalkyl, —C(O)R₇,—(CH₂)_(m)C(O)OR₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁, —S(O)_(p)R₇,—S(O)_(p)OR₇, or —S(O)_(p)NR₁₀OR₁₁; R₄₃ and R₄₄ are, independently, —H,—OH, an optionally substituted alkyl, an optionally substituted alkenyl,an optionally substituted alkynyl, an optionally substituted cycloalkyl,an optionally substituted cycloalkenyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, an optionally substitutedheteraralkyl, hydroxyalkyl, alkoxyalkyl, halo, cyano, nitro, guanadino,a haloalkyl, a heteroalkyl, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁,—NR₈C(O)R₇, —SR₇, —S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇,—NR₈S(O)_(p)R₇, —S(O)_(p)NR₁₀R₁₁, or R₄₃ and R₄₄ taken together with thecarbon atoms to which they are attached form an optionally substitutedcycloalkenyl, an optionally substituted aryl, an optionally substitutedheterocyclyl, or an optionally substituted heteroaryl; R₄₅ is —H, —OH,—SH, —NR₇H, —OR₂₆, —SR₂₆, —NHR₂₆, —O(CH₂)_(m)OH, —O(CH₂)_(m)SH,—O(CH₂)_(m)NR₇H, —S(CH₂)_(m)OH, —S(CH₂)_(m)SH, —S(CH₂)_(m)NR₇H,—OC(O)NR₁₀R₁₁, —SC(O)NR₁₀R₁₁, —NR₇C(O)NR₁₀R₁₁, —OC(O)R₇, —SC(O)R₇,—NR₇C(O)R₇, —OC(O)OR₇, —SC(O)OR₇, —NR₇C(O)OR₇, —OCH₂C(O)R₇, —SCH₂C(O)R₇,—NR₇CH₂C(O)R₇, —OCH₂C(O)OR₇, —SCH₂C(O)OR₇, —NR₇CH₂C(O)OR₇,—OCH₂C(O)NR₁₀R₁₁, —SCH₂C(O)NR₁₀R₁₁, —NR₇CH₂C(O)NR₁₀R₁₁, —OS(O)_(p)R₇,—SS(O)_(p)R₇, —NR₇S(O)_(p)R₇, —OS(O)_(p)NR₁₀R₁₁, —SS(O)_(p)NR₁₀OR₁₁,—NR₇S(O)_(p)NR₁₀R₁₁, —OS(O)_(p)OR₇, —SS(O)_(p)OR₇, —NR₇S(O)_(p)OR₇,—OC(S)R₇, —SC(S)R₇, —NR₇C(S)R₇, —OC(S)OR₇, —SC(S)OR₇, —NR₇C(S)OR₇,—OC(S)NR₁₀OR₁₁, —SC(S)NR₁₀OR₁₁, —NR₇C(S)NR₁₀R₁₁, —OC(NR₈)R₇, —SC(NR₈)R₇,—NR₇C(NR₈)R₇, —OC(NR₈)OR₇, —SC(NR₈)OR₇, —NR₇C(NR₈)OR₇, —OC(NR₈)NR₁₀R₁₁,—SC(NR₈)NR₁₀R₁₁, or —NR₇C(NR₈)NR₁₀OR₁₁; and R₄₆, for each occurrence, isindependently selected from the group consisting of H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, an optionally substituted heteraralkyl,halo, cyano, nitro, guanadino, a haloalkyl, a heteroalkyl, —NR₁₀R₁₁,—OR₇, —C(O)R₇, —C(O)OR₇, —OC(O)R₇, —C(O)NR₁₀R₁₁, —NR₈C(O)R₇, —SR₇,—S(O)_(p)R₇, —OS(O)_(p)R₇, —S(O)_(p)OR₇, —NR₈S(O)_(p)R₇, or—S(O)_(p)NR₁₀R₁₁.
 68. The compound of claim 67, wherein X₄₁ is NR₄₂ andX₄₂ is CR₄₄; R₄₁ is selected from the group consisting of methyl, ethyl,propyl, isopropyl, cyclopropyl, methoxy, ethoxy, propoxy, andcyclopropoxy; and R₄₂ is selected from the group consisting of —H alower alkyl, a lower cycloalkyl, —C(O)N(R₂₇)₂, and —C(O)OH wherein eachR₂₇ is independently —H or a lower alkyl. 69-72. (canceled)
 73. Thecompound of claim 67, wherein X_(4i) is NR₄₂, and R₄₂ is selected fromthe group consisting of —H, methyl, ethyl, n-propyl, isopropyl,cyclopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, —C(O)OH,—(CH₂)_(m)C(O)OH, —CH₂OCH₃, —CH₂CH₂OCH₃, and —C(O)N(CH₃)₂.
 74. Thecompound of claim 67, wherein R₄₃ and R₄₄ are, independently, selectedfrom the group consisting of —H, methyl, ethyl, propyl, isopropyl,cyclopropyl, methoxy, ethoxy, propoxy, and cyclopropoxy; or R₄₃ and R₄₄are taken together with the carbon atoms to which they are attached forma C₅-C₈ cycloalkenyl or a C₅-C₃ aryl. 75-76. (canceled)
 77. The compoundof claim 67, wherein R₄₅ is selected from the group consisting of —H,—OH, —SH, —NH₂, a lower alkoxy, a lower alkyl amino, and a lower dialkylamino.
 78. The compound of claim 77, wherein R₄₅ is selected from thegroup consisting of —H, —OH, methoxy and ethoxy.
 79. (canceled)
 80. Thecompound of claim 67, represented by the following structural formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or clathratethereof.
 81. The compound of claim 80, wherein X₄₂ is CR₄₄, and R₄₃ andR₄₄ are, independently, selected from the group consisting of —H,methyl, ethyl, propyl, isopropyl, cyclopropyl, methoxy, ethoxy, propoxy,and cyclopropoxy; or R₄₃ and R₄₄, are taken together with the carbonatoms to which they are attached, form a C₅-C₈ cycloalkenyl or a C₅-C₈aryl. 82-86. (canceled)
 87. The compound of claim 67, represented by thefollowing structural formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or clathratethereof; wherein: R₄₁ is selected from the group consisting of loweralkyl, lower alkoxy, lower cycloalkyl, and lower cycloalkoxy; and R₄₂ is—H or an optionally substituted lower alkyl.
 88. (canceled)
 89. Thecompound of claim 87, wherein R₄₁ is selected from the group consistingof methyl, ethyl, propyl, isopropyl, cyclopropyl, methoxy, ethoxy,propoxy, and cyclopropoxy.
 90. (canceled)
 91. The compound of claim 1,wherein the compound is selected from the group consisting of4-(4-(2,3-dihydro-1H-inden-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyldihydrogen phosphate;5-hydroxy-4-(5-hydroxy-4-(6-morpholinopyridin-3-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyldihydrogen phosphate;5-hydroxy-4-(5-hydroxy-4-(1-methyl-1H-indol-5-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyldihydrogen phosphate; sodium5-hydroxy-4-(5-hydroxy-4-(1-methyl-1H-indol-5-yl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenylphosphate;2-(4-(2,3-dihydro-1H-inden-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-4-isopropylphenyldihydrogen phosphate;4-(2,3-dihydro-1H-inden-5-yl)-5-(2,4-dihydroxy-5-isopropylphenyl)-4H-1,2,4-triazol-3-yldihydrogen phosphate;4-(4-1′,3′-dihydrospiro[[1,3]dioxolane-2,2′-indene]-5′-yl)-5-mercapto-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyldihydrogen phosphate;2-(3,4-dimethoxyphenethyl)-5-hydroxy-4-(5-hydroxy-4-(1-methyl-1H-indol-5-yl)-4H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate;4-(4-(2,3-dihydro-1H-inden-5-yl)-5-(phenylamino)-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyldihydrogen phosphate;5-hydroxy-2-isopropyl-4-(5-mercapto-4-(4-methoxybenzyl)-4H-1,2,4-triazol-3-yl)phenyldihydrogen phosphate;5-hydroxy-4-(5-hydroxy-4-(4-methoxybenzyl)-4H-1,2,4-triazol-3-yl)-2-isopropylphenyldihydrogen phosphate;4-(4-((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)methyl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-5-hydroxy-2-isopropylphenyldihydrogen phosphate;4-(4-(4-bromo-2-methylphenyl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-3-hydroxyphenyldihydrogen phosphate; or4-(4-(1,3-dimethyl-1H-indol-5-yl)-5-hydroxy-4H-1,2,4-triazol-3-yl)-2-ethyl-5-hydroxyphenyldihydrogen phosphate; or a tautomer, pharmaceutically acceptable salt,solvate, or clathrate, or a prodrug thereof.
 92. A method of inhibitingHsp90 or modulating the activity of glucocorticoid receptors in a cell,comprising administering to the cell an effective amount of a compoundof any one of claim
 1. 93. A method of treating or preventingproliferative disorder or treating cancer in a mammal, comprisingadministering to the mammal an effective amount of a compound ofclaim
 1. 94. (canceled)
 95. A method of inducing degradation of a c-kitprotein, Bcr-Abl protein, a FLT3 protein, an EGFR protein, comprisingadministering to the mammal an effective amount of a compound ofclaim
 1. 96. A method of treating a c-kit associated cancer, Bcr-Ablassociated cancer, FLT3 associated cancer, or an EGFR associated in amammal, comprising administering to the mammal an effective amount of acompound of claim
 1. 97-102. (canceled)
 103. A method of treating orinhibiting angiogenesis, treating a non-Hodgkin's lymphoma, treating orpreventing an inflammatory disorder, inhibiting topoisomerase II,suppressing the immune system, or treating or preventing an immunedisorder in a subject in need thereof, comprising administering to thesubject an effective amount of a compound of claim
 1. 104. A method ofblocking, occluding, or otherwise disrupting blood flow inneovasculature, comprising contacting the neovasculature with aneffective amount of a compound of claim
 1. 105-112. (canceled)
 113. Amethod of treating or preventing a fungal infection, a bacterialinfection, a viral infection, or a parasitic infection in a subject,comprising administering to the subject an effective amount of acompound of claim
 1. 114-122. (canceled)
 123. A pharmaceuticalcomposition, comprising a pharmaceutically acceptable carrier and acompound of claim
 1. 124-134. (canceled)